U.S. patent application number 12/024490 was filed with the patent office on 2009-08-06 for transmitting/reflecting emanating light with time variation.
Invention is credited to Michael Bassler, Markus Beck, Tobias Buergel, Alex Hegyi, Noble M. Johnson, Peter Kiesel.
Application Number | 20090195773 12/024490 |
Document ID | / |
Family ID | 40931335 |
Filed Date | 2009-08-06 |
United States Patent
Application |
20090195773 |
Kind Code |
A1 |
Bassler; Michael ; et
al. |
August 6, 2009 |
Transmitting/Reflecting Emanating Light With Time Variation
Abstract
A filter arrangement can transmit and/or reflect light emanating
from a moving object so that the emanating light has time
variation, and the time variation can include information about the
object, such as its type. For example, emanating light from
segments of a path can be transmitted/reflected through positions
of a filter assembly, and the transmission functions of the
positions can be sufficiently different that time variation occurs
in the emanating light between segments. Or emanating light from a
segment can be transmitted/reflected through a filter component in
which simpler transmission functions are superimposed, so that time
variation occurs in the emanating light in accordance with
superposition of two simpler non-uniform transmission functions.
Many filter arrangements could be used, e.g. the filter component
could include the filter assembly, which can have one of the
simpler non-uniform transmission functions. Time-varying waveforms
from sensing results can be compared to obtain spectral
differences.
Inventors: |
Bassler; Michael; (Menlo,
CA) ; Beck; Markus; (Palo Alto, CA) ; Kiesel;
Peter; (Palo Alto, CA) ; Hegyi; Alex; (Ann
Arbor, MI) ; Buergel; Tobias; (Braunschweig, DE)
; Johnson; Noble M.; (Menlo Park, CA) |
Correspondence
Address: |
Leading Edge Law Group, PLC/Xerox-PARC
Three James Center, 1051 East Cary Street, Suite 1130
Richmond
VA
23219
US
|
Family ID: |
40931335 |
Appl. No.: |
12/024490 |
Filed: |
February 1, 2008 |
Current U.S.
Class: |
356/73 ;
359/890 |
Current CPC
Class: |
G01N 21/255 20130101;
G01N 2021/0346 20130101; G01N 21/05 20130101; G01N 21/645
20130101 |
Class at
Publication: |
356/73 ;
359/890 |
International
Class: |
G02B 5/22 20060101
G02B005/22; G01N 21/00 20060101 G01N021/00 |
Claims
1. A method of using a filter arrangement, the method comprising:
while an object travels along a path past the filter arrangement
and emanates light within an application's range of photon
energies, transmitting/reflecting at least some of the emanating
light through the filter arrangement; the act of
transmitting/reflecting at least some of the emanating light
through the filter arrangement including at least one of: while the
object is in each of two or more segments of the path,
transmitting/reflecting respective portions of the emanating light
through respective positions of a filter assembly within the filter
arrangement; each of the respective positions having a respective
transmission function within the range; the transmission functions
of at least two of the positions being sufficiently different that
time variation occurs in the emanating light between at least two
of the segments; and while the object is in each of a series of
segments of the path, transmitting/reflecting a respective portion
of the emanating light through a filter component within the filter
arrangement; within the range, the filter component having a
combined transmission function in which a set of two or more
simpler transmission functions is superimposed; the set including
first and second simpler non-uniform transmission functions, the
set being superimposed such that time variation occurs in the
emanating light in accordance with superposition of the first and
second simpler non-uniform transmission functions.
2. The method of claim 26 in which the act of
transmitting/reflecting at least some of the emanating light
through the filter arrangement includes both the act of
transmitting/reflecting respective portions of the emanating light
through a longitudinal sequence of two or more filter elements and
also the act of transmitting/reflecting a respective portion of the
emanating light through a filter component.
3. The method of claim 1 in which the filter assembly includes
first and second filter elements that include first and second ones
of the respective positions, respectively; each of the first and
second filter elements having a respective transmission function
that is approximately uniform for light from the respective
segment.
4. The method of claim 3 in which the transmission functions of the
first and second filter elements are spectrally different; the act
of transmitting/reflecting respective portions of the emanating
light through respective positions including: encoding spectral
information in time variation of the emanating light.
5. The method of claim 3 in which the transmission functions of the
first and second filter elements are different in
transmitted/reflected intensity; the act of transmitting/reflecting
respective portions of the emanating light through respective
positions including: encoding intensity information in time
variation of the emanating light.
6. The method of claim 26 in which at least one of the simpler
transmission functions is non-periodic.
7. The method of claim 26, further comprising: causing the object
to travel along the path with non-uniform displacement.
8. Apparatus comprising: a fluidic structure that includes a
channel through which objects can travel along respective paths
during operation of the apparatus; and an encoding component; the
encoding component including a filter arrangement that can receive
light emanating from objects in the channel; in response to input
light that is within an application's range of photon energies and
that is emanating from an object traveling through the channel past
the filter arrangement, the filter arrangement providing output
light; the filter arrangement including at least one of: a filter
assembly that includes a set of positions, each having a respective
transmission function within the range; a sequence of segments of
an object's path including at least two segments from which
respective positions in the set receive light emanating from the
object, the transmission functions of at least two of the
respective positions being sufficiently different from each other
that time variation occurs in the output light while the object
travels through the sequence of segments; and a filter component
that receives input light from a segment of an object's path and
that, within the range, has a combined transmission function in
which a set of two or more simpler transmission functions is
superimposed; the set including first and second simpler
non-uniform transmission functions, the set being superimposed such
that time variation occurs in the output light in accordance with
superposition of the first and second simpler non-uniform
transmission functions while the object travels through the
segment.
9. The apparatus of claim 27 in which the filter component includes
the longitudinal sequence, the longitudinal sequence having one of
the first and second simpler non-uniform transmission
functions.
10. The apparatus of claim 27, further comprising: a photosensing
component that photosenses the output light and provides sensing
results; and a processing component that, in response to the
sensing results, performs operations to obtain data indicating
information encoded in the output light.
11. The apparatus of claim 10 in which the sensing results indicate
one or more sensed time-varying waveforms, the processing component
being programmed to: perform a comparing operation on a set of
time-varying waveforms to obtain comparison results, at least one
of the set being a sensed time-varying waveform; and use the
comparison results to obtain data indicating at least one spectral
difference between the time-varying waveforms in the set.
12. The apparatus of claim 8 in which the filter arrangement
includes at least one of: an absorption filter; an
interference-based filter; a light-transmissive and/or
light-reflective filter; a longitudinal sequence of filters that
vary in a periodic, random, or chirped pattern; filter elements of
two or more colors; filter elements of two or more gray levels;
overlapping filter elements; a longitudinal sequence of filter
elements that includes binary, gray level, and color filter
elements; two or more lengthwise extending filter elements; a
radial sequence of filter elements; two orthogonally striped filter
assemblies; filter assemblies on opposite sides of a fluidic
channel; and a stack-equivalent filter.
13. The apparatus of claim 27 in which at least one of the simpler
non-uniform transmission functions is a periodic, random, or chirp
function of position.
14. The apparatus of claim 27 in which the objects are biological
cells or viruses and the apparatus is a flow cytometer.
15. A method comprising: while objects travel along respective
paths past a filter arrangement, transmitting/reflecting light
emanating from the objects through the filter arrangement, the
emanating light including photon energies within a range of photon
energies; the act of transmitting/reflecting light through the
filter arrangement including: while an object travels through a
segment of its respective path, transmitting/reflecting at least
some of the object's emanating light through a longitudinal
sequence of two or more filter elements within the filter
arrangement; the longitudinal sequence including a first subset of
one or more filter elements each having approximately a first
transmission function within the range of photon energies and a
second subset of one or more filter elements each having
approximately a second transmission function within the range of
photon energies, the first and second transmission functions being
sufficiently spectrally different from each other that time
variation occurs in the emanating light while the object travels
through the segment.
16. The method of claim 15 in which the time variation encodes
spectral information about the object, the method further
comprising: photosensing the emanating light to obtain sensing
results; and using the sensing results to obtain data indicating
the encoded spectral information.
17. The method of claim 16 in which the sensing results indicate
one or more sensed time-varying waveforms; the act of using the
sensing results including: performing a comparing operation on a
set of time-varying waveforms to obtain comparison results, at
least one of the set being a sensed time-varying waveform; and
using the comparison results to obtain data indicating at least one
spectral difference between the time-varying waveforms in the
set.
18. A method comprising: while objects travel along respective
paths past a filter arrangement, transmitting/reflecting light
emanating from the objects through the filter arrangement, the
emanating light including photon energies within a range of photon
energies; the act of transmitting/reflecting light through the
filter arrangement including: while an object travels through a
segment of its path, transmitting/reflecting at least some of the
object's emanating light through a filter component within the
filter arrangement; the filter component having a combined
transmission function in which a set of two or more simpler
transmission functions is superimposed; the set including first and
second simpler non-uniform transmission functions, the set being
superimposed such that time variation occurs in the object's
emanating light in accordance with superposition of the first and
second simpler non-uniform transmission functions while the object
travels through the segment.
19. The method of claim 18 in which at least one of the first and
second simpler non-uniform transmission functions is a
spectrally-dependent transmission function that transmits more than
one color as a function of position.
20. The method of claim 18 in which the first simpler non-uniform
transmission function is a non-periodic spectrally-dependent
transmission function; the second simpler non-uniform transmission
function being a periodic transmission function.
21. The method of claim 18 in which at least one of the first and
second simpler non-uniform transmission functions is a
spectrally-independent transmission function that transmits black
and white and/or gray scales.
22. The method of claim 18 in which at least one of the first and
second simpler non-uniform transmission functions is a periodic,
random, or chirp function of position along the segment.
23. A method of using a photosensor, the method comprising: while a
light-emanating object travels along a path past the photosensor,
photosensing the object's emanating light to obtain sensing
results; and using the sensing results to obtain information about
the object; the emanating light having time variation indicating
spectral information about the object's emission spectrum; the
sensing results indicating one or more sensed time-varying
waveforms; the act of using the sensing results comprising:
performing a comparing operation on a set of time-varying waveforms
to obtain comparison results, at least one of the set being a
sensed time-varying waveform; and using the comparison results to
obtain data indicating the spectral information about the
object.
24. The method of claim 23 in which the act of performing a
comparing operation includes at least one of: performing a
comparing operation that compares two sensed time-varying waveform;
performing a comparing operation that compares a sensed
time-varying waveform and a template; performing comparing
operations, each comparing a sensed time-varying waveform and one
of a number of templates, each template being for a respective type
of object; performing a comparing operation on time-varying
waveforms, one of which has been time scaled; performing an
operation that obtains a similarity measure as a function of
time-alignment; performing a correlation operation and obtaining
comparison results indicating that time-varying waveforms are
correlated; performing a correlation operation and obtaining
comparison results indicating that time-varying waveforms are
anti-correlated; and performing correlation on a continuous basis
and continuously obtaining comparison results.
25. The apparatus of claim 23 in which act of using the comparison
results includes at least one of: performing an operation that
obtains a time derivative of a waveform indicating comparison
results; performing an operation that obtains a derivative waveform
with a positive peak followed by a negative peak; performing an
operation that obtains a derivative waveform with a negative peak
followed by a positive peak; performing an operation that obtains
an amplitude of a peak in a waveform indicating comparison results;
performing an operation that obtains an amplitude between positive
and negative peaks in a derivative waveform; and obtaining data
indicating spectral difference.
26. A method of using filter arrangements, the method comprising:
while an object travels along a path past a filter arrangement and
emanates light within an application's range of photon energies,
transmitting/reflecting at least some of the emanating light
through the filter arrangement; the act of transmitting/reflecting
at least some of the emanating light through the filter arrangement
including at least one of: while the object travels through a
segment of the path, transmitting/reflecting respective portions of
the emanating light through a longitudinal sequence of two or more
filter elements within the filter arrangement; the longitudinal
sequence including a first subset of one or more filter elements
each having approximately a first transmission function within the
range of photon energies and a second subset of one or more filter
elements each having approximately a second transmission function
within the range of photon energies, the first and second
transmission functions being sufficiently different from each other
that time variation occurs in the emanating light in accordance
with the longitudinal sequence; and while the object is in each of
a series of segments of the path, transmitting/reflecting a
respective portion of the emanating light through a filter
component within the filter arrangement; within the range, the
filter component having a combined transmission function in which a
set of two or more simpler transmission functions are superimposed;
the set including first and second simpler non-uniform transmission
functions, the set being superimposed such that time variation
occurs in the emanating light in accordance with superposition of
the first and second simpler non-uniform transmission
functions.
27. Apparatus comprising: a fluidic structure that includes a
channel through which objects can travel along respective paths
during operation of the apparatus; and an encoding component; the
encoding component including a filter arrangement that can receive
light emanating from objects in the channel; in response to input
light that is within an application's range of photon energies and
that is emanating from an object traveling through the channel past
the filter arrangement, the filter arrangement providing output
light; the filter arrangement including at least one of: a
longitudinal sequence of two or more filter elements; the
longitudinal sequence including a first subset of one or more
filter elements each having approximately a first transmission
function within the range of photon energies and a second subset of
one or more filter elements each having approximately a second
transmission function within the range of photon energies, the
first and second transmission functions being sufficiently
different from each other that time variation occurs in the output
light in accordance with the longitudinal sequence; and a filter
component that receives input light from a segment of an object's
path and that, within the range, has a combined transmission
function in which a set of two or more simpler transmission
functions is superimposed; the set including first and second
simpler non-uniform transmission functions, the set being
superimposed such that time variation occurs in the output light in
accordance with superposition of the first and second simpler
non-uniform transmission functions while the object travels through
the segment.
28. The apparatus of claim 27, further comprising: a photosensing
component that photosenses the output light; the apparatus further
comprising one of: a light-transmissive component that bounds the
channel and that is between the channel and the photosensing
component, the filter arrangement being at a surface of the
light-transmissive component toward the channel; the apparatus
being without an optical component between the light-transmissive
component and the photosensing component; a photosensitive surface
of the photosensing component bounding the channel, the filter
arrangement being on the photosensitive surface; the apparatus
being without an optical component between the channel and the
filter arrangement; and a light-transmissive component that bounds
the channel and that is between the channel and the photosensing
component, the filter arrangement being on a photosensitive surface
of the photosensing component toward the light-transmissive
component; the apparatus being without an optical component between
the light-transmissive component and the filter arrangement.
29. The apparatus of claim 27 in which the photosensing component
includes at least one of: a large area photosensor; an array of
photosensing cells; and an array of discrete photosensors.
Description
[0001] The following applications, each of which is hereby
incorporated by reference in its entirety, might be regarded as
related to this application: "Additive Printed Mask Process and
Structures Produced Thereby", U.S. patent application Ser. No.
10/536,102, now published as U.S. Patent Publication No.
2007/0172969; "Sensing Photon Energies Emanating from Channels or
Moving Objects", U.S. patent application Ser. No. 11/315,386, now
published as U.S. Patent Publication No. 2007/0146704; "Sensing
Photon Energies of Optical Signals", U.S. patent application Ser.
No. 11/315,926, now published as U.S. Patent Publication No.
2007/0147189; "Sensing Photons from Objects in Channels", U.S.
patent application Ser. No. 11/315,992, now published as U.S.
Patent Publication No. 2007/0145249; "Obtaining Analyte
Information", U.S. patent application Ser. No. 11/316,303, now
published as U.S. Patent Publication No. 2007/0148760; "Providing
Light to Channels or Portions", U.S. patent application Ser. No.
11/316,660, now published as U.S. Patent Publication No.
2007/0147728; "Method and System for Evaluation of Signals Received
from Spatially Modulated Excitation and Emission to Accurately
Determine Particle Positions and Distances", U.S. patent
application Ser. No. 11/698,338; "Method and System Implementing
Spatially Modulated Excitation or Emission for Particle
Characterization with Enhanced Sensitivity", U.S. patent
application Ser. No. 11/698,409; "Obtaining Information From
Optical Cavity Output Light", U.S. patent application Ser. No.
11/702,249; "Photosensing Optical Cavity Output Light", U.S. patent
application Ser. No. 11/702,250; "Distinguishing Objects", U.S.
patent application Ser. No. 11/702,328; "Encoding Optical Cavity
Output Light", U.S. patent application Ser. No. 11/702,363; "Moving
Analytes and Photosensors", U.S. patent application Ser. No.
11/702,470; "Surface Energy Control Methods for Color Filter
Printing", U.S. patent application Ser. No. 11/755,717; "Producing
Sandwich Waveguides", U.S. patent application Ser. No. 11/777,661;
"Producing Fluidic Waveguides", U.S. patent application Ser. No.
11/777,712; "Obtaining Information from Time Variation of Sensing
Results", U.S. patent application Ser. No. 12/022,485; and
"Providing Time Variation in Emanating Light", U.S. patent
application Ser. No. 12/023,436.
BACKGROUND OF THE INVENTION
[0002] The present invention relates generally to techniques that
transmit and/or reflect light emanating from objects. More
specifically, techniques can use filter arrangements to transmit
and/or reflect such light with time variation, such as where the
objects are moving relative to the filter arrangements.
[0003] Various techniques have been proposed for using light
emanating from objects. For example, U.S. Patent Application
Publication No. 2007/0145249 describes a fluidic structure with a
channel along which is a series of sensing components to obtain
information about objects traveling within the channel, such as
droplets or other objects carried by fluid. A sensing component
includes a set of cells that photosense a range of photon energies
that emanate from objects. A processor can receive information
about objects from the sensing components and use it to obtain
spectral information. Similar techniques are described, for
example, in U.S. Patent Application Publication Nos. 2007/016704,
2007/0147189, and 2007/0147728.
[0004] Also, various flow cytometry techniques have been
proposed.
[0005] It would be advantageous to have improved techniques for
using light emanating from objects, including improved techniques
for transmitting and/or reflecting such light with time
variation.
SUMMARY OF THE INVENTION
[0006] The invention provides various exemplary embodiments,
including methods and apparatus. In general, the embodiments
involve transmitting and/or reflecting emanating light through
filter arrangements to obtain time variation.
[0007] These and other features and advantages of exemplary
embodiments of the invention are described below with reference to
the accompanying drawings.
BRIEF DESCRIPTION OF THE DRAWINGS
[0008] FIG. 1 is a schematic diagram showing features of techniques
in which a filter arrangement transmits and/or reflects light
emanating from an object with time variation.
[0009] FIG. 2 is a schematic diagram showing components of a system
in which light emanating from an object can include information
about characteristics of the object.
[0010] FIG. 3 is a schematic diagram of an excitation arrangement
in an encoding component as in FIG. 2.
[0011] FIG. 4 is a schematic diagram of a filter arrangement in an
encoding component as in FIG. 2.
[0012] FIG. 5 is a schematic diagram of a displacement control
arrangement in an encoding component as in FIG. 2.
[0013] FIG. 6 is a schematic block diagram of a system in which
components as in FIG. 2 can be implemented.
[0014] FIG. 7 is a flow chart showing general operations in an
implementation of an object distinguishing routine as in FIG.
6.
[0015] FIG. 8 is a schematic diagram of an analyzer in a fluidic
structure, where the analyzer includes a system that can be
implemented as in FIGS. 6 and 7.
[0016] FIG. 9 is a top view of an article that can include a filter
arrangement and that can be included in an encoding component as in
FIG. 2.
[0017] FIG. 10 is a cross-sectional view of an implementation of an
article similar to that in FIG. 9, taken along the line 10-10.
[0018] FIG. 11 is a cross-sectional view of another implementation
of an article similar to that in FIG. 9, taken along the line
11-11, together with graphs of sensed intensities.
[0019] FIG. 12 is a partially schematic cross-sectional view
showing two ways in which a filter arrangement on a photosensitive
surface can be configured in an encoding component as in FIG.
2.
[0020] FIG. 13 is a schematic top view of another filter
arrangement that can be included in an encoding component as in
FIG. 2.
[0021] FIG. 14 is a cross-sectional view of an implementation of a
filter arrangement similar to that in FIG. 14, taken along the line
14-14, together with graphs of transmitted intensities.
[0022] FIG. 15 is a cross-sectional view of another implementation
of a filter arrangement that can be included in an encoding
component as in FIG. 2, together with graphs showing spectra of
transmitted intensities.
[0023] FIG. 16 is a cross-sectional view of yet another
implementation of a filter assembly that can be included in an
encoding component as in FIG. 2.
[0024] FIG. 17 is a cross-sectional view of yet another
implementation of a filter assembly that can be included in an
encoding component as in FIG. 2.
[0025] FIG. 18 is a cross-sectional view of yet another
implementation of a filter assembly that can be included in an
encoding component as in FIG. 2, such as with features as in any of
FIGS. 15-17.
[0026] FIG. 19 is another partially schematic cross-sectional view
showing a displacement control arrangement that includes shaped
boundaries, together with graphs showing velocity of an object and
also showing intensity of emanating light as a function of
time.
[0027] FIG. 20 is a cross-sectional view of another displacement
control arrangement that can be included in an encoding component
as in FIG. 2, together with graphs showing intensity of emanating
light for exemplary types of objects.
[0028] FIG. 21 is a partially schematic cross-sectional view of
another displacement control arrangement that can be included in an
encoding component as in FIG. 2, together with a graph showing
displacement as a function of time and graphs showing intensity of
emanating light as a function of time for exemplary types of
objects.
[0029] FIG. 22 is a top view of an implementation of a fluidic
channel with an encoding arrangement that can be included in an
implementation with features as in FIG. 1.
[0030] FIG. 23 is a cross-sectional view of a component in FIG. 22,
taken along the line 23-23.
[0031] FIG. 24 is a cross-sectional view of another component in
FIG. 22, taken along the line 24-24.
[0032] FIG. 25 includes a set of graphs showing cross-sectional
thickness as a function of position in an x-direction for filters
and showing transmission as a function of position in the
x-direction or as a function of time t for one of the filters.
[0033] FIG. 26 includes a set of graphs showing cross-sectional
thickness as a function of position in an x-direction for other
filters and showing transmission as a function of position in the
x-direction or as a function of time t for one of the filters.
[0034] FIG. 27 is a schematic cross-sectional view of a filter
assembly that includes two simpler filters.
[0035] FIG. 28 is a flow chart with graphs illustrating an
implementation in which information about objects is obtained from
sensed time-varying signals.
DETAILED DESCRIPTION
[0036] In the following detailed description, numeric values and
ranges are provided for various aspects of the implementations
described. These values and ranges are to be treated as examples
only, and are not intended to limit the scope of the claims. In
addition, a number of materials are identified as suitable for
various facets of the implementations. These materials are to be
treated as exemplary, and are not intended to limit the scope of
the claims.
[0037] "Light" refers herein to electromagnetic radiation of any
wavelength or frequency; unless otherwise indicated, a specific
value for light wavelength or frequency is that of light
propagating through vacuum. Light that can include information is
sometimes referred to herein as an "optical signal".
[0038] The term "sensing" is used herein in the most generic sense
of obtaining information from a physical stimulus; sensing
therefore includes actions such as detecting, measuring, and so
forth. A "sensor" is a device that performs sensing. Data or other
signals that indicate or include results of sensing are sometimes
referred to herein as "sensing results".
[0039] "Photosensing" is sensing of light. A "photosensor" is
accordingly an electronic device that performs photosensing. More
specifically, if optical signals include information, a photosensor
that receives the optical signals may be able to sense the
information and provide sensing results that indicate or include
the information. A surface at which photosensing occurs is referred
to herein as a "photosensitive surface".
[0040] The various exemplary implementations described below
address problems that arise in obtaining information about a moving
object such as a biological cell, a virus, a molecule, or a
submolecular complex, such as in flow cytometry. Flow cytometry has
become an indispensable tool in clinical diagnostics, such as in
diagnosing cancer, AIDS, and infectious diseases during outbreaks,
and also in microbiology and other areas. The cost and size of
existing cytometers preclude their use in field clinics, water
monitoring, agriculture/veterinary diagnostics, and rapidly
deployable biothreat detection.
[0041] A number of commercially available flow cytometers use
multiple excitation sources, each focused on a well-defined
location or region separate from the others. Light emitted from
each source's region is typically analyzed with a series of beam
splitters, filters, and photomultiplier tubes (PMTs) in order to
detect and distinguish differently stained cells or cells that
concurrently carry multiple dyes. Cells are typically stained in
solution with different dyes prior to insertion into a cytometer,
and the measurement takes place in a fluidic channel in which cells
traverse a detection region, typically at a speed of up to several
meters per second. In the detection region, focused laser light
(typically with an elliptical focus of 80 .mu.m.times.40 .mu.m)
excites the dyes on the cells. The resulting fluorescent light can
be collected by a microscope lens, sorted by band pass filters, and
detected by PMTs or avalanche photodiodes (APDs). For each spot
excitation, a respective set of filters and detectors is needed,
which is costly and leads to bulky devices with strict requirements
necessary to maintain optical alignment. Since the detection region
is small and objects traverse it rapidly (typical dwell times are
around 10 .mu.sec), such flow cytometers have serious
signal-to-noise (S/N) ratio issues for weakly fluorescing cells.
These issues become more acute if multiple targets must be
characterized and distinguished in some way, such as by
counting.
[0042] A major cost in flow cytometry applied in clinical
diagnostics is cost of reagents (e.g. antibodies and conjugated
dyes). There are two ways to reduce the amount of consumables:
First, one can reduce the required amount of analyte, e.g. by
employing microfluidic techniques; and second, one can reduce the
amount of consumable per analyte volume. Reducing amounts used
would, however, reduce fluorescent intensity. It would be valuable
to be able to overcome this constraint with a cost-effective and
reliable technique to detect and distinguish weakly emitting
cells.
[0043] Previous proposals to address these problems have involved
spatially modulated single-color excitation to improve S/N ratios
and to shift the detection limit toward weaker emitting cells.
Spatial resolution can be maintained or improved in comparison with
previous flow cytometry techniques, because fluorescing light is
spatially modulated over a comparably large detection region; this
is helpful because spatial resolution affects maximum detection or
count rate of a device. But single-color techniques are limited,
whether excitation is performed in a black/white approach or with a
single-color interference pattern from a light source. Also,
single-color techniques can encounter problems with wavelength
sensitivity and bleaching of dyes. Because of low wavelength
sensitivity, many flow cytometers with filter-PMT combinations are
also constrained to use dyes with substantially different
fluorescence wavelengths.
[0044] In addressing such problems, some exemplary implementations
described below employ filter arrangements that transmit or reflect
emanating light with one or both of two techniques: A filter
assembly is used that provides different transmission functions in
different segments of an object's path and/or a filter component is
used that has a combined transmission function in which a set of
simpler non-uniform transmission functions are superimposed. Such
techniques make it possible to provide several different
transmission functions in sequence within relatively short part of
an object's path, so that the object's emanating light is
relatively constant across the different transmission functions.
These techniques also allow much greater variation in filter
arrangements than would be possible with binary, black/white masks
or single color masks. In addition, these techniques can be
implemented to maintain higher spatial resolution and to allow
higher photon flux on a photosensor. Time variation of emanating
light resulting from such filters may provide sufficient
information to make spectral characterization of particles
feasible. Use of multiple colors may be compatible with particle
identification based on native fluorescence; in particular,
patterned filter arrangements allow for detection of differences in
emission spectra and even the very small differences that occur in
native fluorescence spectra might be detectable. It may also enable
advanced color monitoring in printing applications by detecting
even small differences in the reflection spectra of color spots
while they are moving past interdigitated or otherwise patchworked
or patterned filter arrangements.
[0045] The term "photon" refers herein to a quantum of light, and
the term "photon energy" refers herein to the energy of a photon.
Light can be described as having a "photon energy distribution" or,
more commonly, a "spectrum", meaning the combination of photon
energies that are included in the light; highly monochromatic
light, for example, has a photon energy distribution or spectrum
with one peak energy value.
[0046] Light can also be described as provided by a "light source,"
which, unless otherwise specified, refers herein to any device,
component, or structure that can provide light of the type
described; examples of light sources relevant to the
below-described implementations include various kinds of pulsed and
unpulsed lasers and laser structures, laser diodes (LDs), light
emitting diodes (LEDs), superluminescent LEDs (SLEDs), resonant
cavity LEDs, sources of broadband light that is spectrally filtered
such as with a monochromator, and so forth.
[0047] To "propagate" light through a region or structure is to
transmit or otherwise cause the light to propagate through the
region or structure. The light may be referred to as "propagated
light" or "propagating light".
[0048] Propagating light can often be usefully characterized by
direction and speed of propagation, with direction typically
illustrated by one or more rays and with speed typically being
described relative to the constant c, also referred to as the speed
of light in vacuum. Where the speed of light in a medium M is a
constant c.sub.M less than c, then M has an index of refraction
n.sub.M=c/c.sub.M.
[0049] Where light changes direction in a way that can be
illustrated or approximated as a vertex between an incoming ray and
an outgoing ray that are both on one side of a surface, the change
may be referred to as a "reflection"; similarly, to "reflect" light
is to cause the light to change its direction of propagation
approximately at such a surface, referred to herein as a
"reflection surface". Similarly, where light changes direction in a
way that can be illustrated or approximated as a vertex between an
incoming ray and an outgoing ray that are on opposite sides of a
surface between two media with different indices of refraction, the
change may be referred to as a "refraction"; similarly, to
"refract" light is to cause the light to change its direction of
propagation approximately at such a surface, referred to herein as
a "refraction surface". In many practical applications, both
reflection and refraction occur at a surface, which may be referred
to herein as a "partially reflecting surface".
[0050] Where light propagates at less than c, it may be useful to
obtain an "optical distance" of propagation; for any segment of
length d in which speed of propagation is constant .epsilon.*c,
where .epsilon.=1/n.sub.EFF.ltoreq.1 and n.sub.EFF is an effective
index of refraction for the segment, optical distance
D(.epsilon.)=d/.epsilon.. An optical distance may be referred to
herein as an "optical thickness", such as where light is
propagating through a thickness of material.
[0051] A "range of photon energies" or an "energy range" is a range
of energy values that photons can have. An energy range can be
described, for example, as a range of wavelengths or a range of
frequencies or, in appropriate cases, by the range's central
wavelength or frequency and possibly also the range's width. A
"subrange" of a range of photon energies is a part of the range,
and can be similarly described.
[0052] In general, the upper and lower boundaries and widths of
ranges and subranges are approximate. To provide output photons or
to photosense quantity of photons "throughout", "within", or "in" a
range or subrange means to provide photons or to obtain information
about quantity of photons that are predominantly within the range
or subrange. In typical cases, between 60-90% of the provided
photons or sensed quantity of photons have energies within the
range or subrange, but the percentage could be lower or higher. In
some applications, 90% or even 95% or more of the provided photons
or sensed quantity of photons have energies within the range or
subrange.
[0053] The term "electrical signal" is used herein to encompass any
signal that transfers information from one position or region to
another in an electrical, electronic, electromagnetic, or magnetic
form. Electrical signals may be conducted from one position or
region to another by electrical or magnetic conductors, but the
broad scope of electrical signals also includes light and other
electromagnetic forms of signals and other signals transferred
through non-conductive regions due to electrical, electronic,
electromagnetic, or magnetic effects. In general, the broad
category of electrical signals includes both "analog" and "digital"
signals: An "analog" electrical signal includes information in the
form of a continuously variable physical quantity, such as voltage;
a "digital" electrical signal, in contrast, includes information in
the form of discrete values of a physical characteristic, which
could also be, for example, voltage.
[0054] Some implementations of filter arrangements described herein
employ structures with one or more dimensions smaller than 1 mm,
and various techniques have been proposed for producing such
structures. In particular, some techniques for producing such
structures are referred to as "microfabrication." Examples of
microfabrication include various techniques for depositing
materials such as growth of epitaxial material, sputter deposition,
evaporation techniques, plating techniques, spin coating, printing,
and other such techniques; techniques for patterning materials,
such as etching or otherwise removing exposed regions of thin films
through a photolithographically patterned resist layer or other
patterned layer; techniques for polishing, planarizing, or
otherwise modifying exposed surfaces of materials; and so
forth.
[0055] In the implementations described below, structures, systems,
or parts or components of structures or systems may sometimes be
referred to as "attached" to each other or to other structures,
systems, parts, or components or visa versa, and operations are
performed that "attach" structures, systems, or parts or components
of structures or systems to each other or to other things or visa
versa; the terms "attached", "attach", and related terms refer to
any type of connecting that could be performed in the context. One
type of attaching is "mounting", which occurs when a first part or
component is attached to a second part or component that functions
as a support for the first. In contrast, the more generic term
"connecting" includes not only "attaching" and "mounting", but also
making other types of connections such as electrical connections
between or among devices or components of circuitry. A combination
of one or more parts connected in any way is sometimes referred to
herein as a "structure".
[0056] Some of the structures, elements, and components described
herein are supported on a "support structure" or "support surface",
which terms are used herein to mean a structure or a structure's
surface that can support other structures. More specifically, a
support structure could be a "substrate", used herein to mean a
support structure on a surface of which other structures can be
formed or attached by microfabrication or similar processes.
[0057] The surface of a substrate or other support surface is
treated herein as providing a directional orientation as follows: A
direction away from the surface is "up", "over", or "above", while
a direction toward the surface is "down", "under", or "below". The
terms "upper" and "top" are typically applied to structures,
components, or surfaces disposed away from the surface, while
"lower" or "underlying" are applied to structures, components, or
surfaces disposed toward the surface. In general, it should be
understood that the above directional orientation is arbitrary and
only for ease of description, and that a support structure or
substrate may have any appropriate orientation.
[0058] A structure may be described by its operation, such as a
"support structure" that can operate as a support as described
above; other examples are defined below. In addition, a structure
may be characterized by the nature of its parts or the way in which
they are connected; for example, a "layered structure" is a
structure that includes one or more layers, and the terms "partial
structure" and "substructure" refer to structures that are in turn
parts of other structures.
[0059] Unless the context indicates otherwise, the terms
"circuitry" and "circuit" are used herein to refer to structures in
which one or more electronic components have sufficient electrical
connections to operate together or in a related manner. In some
instances, an item of circuitry can include more than one circuit.
An item of circuitry that includes a "processor" may sometimes be
analyzed into "hardware" and "software" components; in this
context, "software" refers to stored or transmitted data that
controls operation of the processor or that is accessed by the
processor while operating, and "hardware" refers to components that
store, transmit, and operate on the data. The distinction between
"software" and "hardware" is not always clear-cut, however, because
some components share characteristics of both; also, a given
software component can often be replaced by an equivalent hardware
component without significantly changing operation of circuitry,
and a given hardware component can similarly be replaced by
equivalent processor operations controlled by software.
[0060] Circuitry can be described based on its operation or other
characteristics. For example, circuitry that performs control
operations is sometimes referred to herein as "control circuitry"
and circuitry that performs processing operations is sometimes
referred to herein as "processing circuitry".
[0061] In general, sensors, processors, and other such items may be
included in a system in which they are operated automatically or
partially automatically. As used herein, the term "system" refers
to a combination of two or more parts or components that can
perform an operation together. A system may be characterized by its
operation; for example, an "object distinguishing system" is a
system that operates somehow to distinguish objects.
[0062] Within a system, device, or other article, components and
parts may be referred to in a similar manner. One component of an
object distinguishing system, for example, can be described as an
"encoding component", in some cases referred to as an "encoding
arrangement", in either case meaning that the component or
arrangement operates to encode information; similarly, a system can
include an "filter component", in some cases referred to as an
"filter arrangement", in either case meaning that the component or
arrangement operates to perform filtering, as explained in greater
detail below; various other components are described below. In
addition, a component or part may be identified by characteristics
other than its operation.
[0063] In FIG. 1, object 10 is one of series 12 of objects 14
through 16 that travel along respective paths past filter
arrangement 20. The term "path" is used herein in the general sense
of a series of positions and/or configurations that a moving and/or
varying object can have during its motion and/or variation. For
generality, a part of a path is sometimes referred to herein as a
"segment", which could encompass any continuous series of one or
more positions and/or configurations within a path.
[0064] As object 10 travels past arrangement 20, light emanates
from it, such as by emission, scattering (including, e.g.
reflection), or transmission, and a portion of the emanating light
is received by filter arrangement 20, as indicated by arrow 22. In
general, the emanating light includes light within an application's
range of photon energies, meaning that techniques as in FIG. 1 can
be successfully used in a given application, e.g. flow cytometry,
bio-chip readout, or any suitable kind of analyte detection, even
though emanating light might also include photon energies that are
outside the application's range and that might not interact with
filter arrangement 20 in the same way as light in the application's
range.
[0065] The term "object" is used herein in the general sense of any
distinguishable thing about which information can be obtained by a
sensor and included in its sensing results. In some
implementations, sensors can obtain information about objects by
receiving signals from them; for example, signals in the form of
light can emanate from an object, whether through emission (e.g.
radiation, fluorescence, incandescence, chemoluminescence,
bioluminescence, other forms of luminescence, etc.), scattering
(e.g. reflection, deflection, diffraction, refraction, etc.), or
transmission, and can be sensed by a photosensor. The light
"emanates from" or is simply "from" the object, and may be referred
to herein as "emanating light". An object from which light is
emanating may be referred to herein as a "light-emanating object".
In other implementations, sensors can obtain information about
objects in other ways, some of which are mentioned herein.
[0066] Examples of objects that could occur in implementations as
described below include droplets, small volumes of fluid, single
molecules, agglomerated molecules, molecule clusters, cells,
viruses, bacteria, lengthy polymers such as DNA or protein chains,
submolecular complexes such as tags on DNA or protein chains,
microparticles, nanoparticles, beads or other small particles that
can bind and carry specific chemicals or other analytes, emulsions,
any such type of object in an array such as an array of sample
wells, and a distinguishable region of a surface such as a small
area of a sheet of paper or other image-bearing medium; a
distinguishable region, could, for example, be a colored spot. A
droplet or small volume of fluid may, for example, include atoms,
molecules, or other particles that emit light spontaneously or in
response to excitation; a particle could be a "fluorescent
component" of a droplet, fluorescing in response to excitation. Or
a droplet may include particles that absorb light incident on the
droplet, so that the droplet does not reflect or otherwise scatter
the absorbed light; in this case, a particle could be an "absorbent
component" of a droplet. Or a droplet may include particles that
scatter light incident on the droplet in a way that depends on
photon energy, so that the droplet scatters the incident light
correspondingly; in this case, a particle could be a "scattering
component" of a droplet. An analyte (i.e. a chemical species being
investigated) in a droplet or other object can act as a
fluorescent, absorbent, or scattering component. Analyte that is
otherwise homogeneously distributed, for example, can be localized
by binding to carrier beads, resulting in a moving object that
emanates light or provides other signals in a way that depends on
the analyte.
[0067] With respect to a light-emanating object, the expressions
"characteristic of an object" and "emanating light including
information" have related meanings: The term "characteristic"
refers to a trait, quality, or property of an object that can be
measured and that persists with a given value or within a given
range or other subset of possible values while light that "includes
information about the characteristic" is emanating from the object.
In appropriate implementations, characteristics of an object could
include mass, volume, density, cross-section or other shape,
chemical composition, position, speed, acceleration, direction of
movement, spin axis, directional or angular velocity or momentum,
net charge, charge polarity, absorption spectrum, emission
spectrum, scattering spectrum, and so forth. Therefore, emanating
light "includes" information about a characteristic of an object if
information included in the emanating light indicates a value,
range, or other measure of the characteristic. Similar terminology
can apply to types of signals other than emanating light.
[0068] Emanating light or other types of signals can "include
information" in many ways, some of which are described below in
relation to specific implementations. Various criteria could be
used to determine whether emanating light or another type of signal
includes specified information, and such criteria can be referred
to as "encoding criteria". Some encoding criteria, for example,
involve comparison of magnitude of a signal with noise magnitude,
e.g. signal-to-noise (S/N) ratios, because S/N ratio can affect
whether specified information can be recovered from sensing results
obtained by photosensing emanating light. Other types of encoding
criteria could be used as appropriate. Where emanating light or
another type of signal satisfies an appropriate encoding criterion
for specified information, the light or signal may be said to
"encode" the information.
[0069] Similarly, sensing results, whether from photosensing
emanating light or from another type of sensing, can "include
information" in many ways, and similar encoding criteria could be
applied as with signals. Where sensing results indicate one or more
time-varying waveforms, the sensing results can be referred to as
having "encoded time variation".
[0070] The term "waveform" is used herein in the general sense of
any set of values that varies over one or more dimensions, whether
continuous or discrete, whether analog or digital, and whether
measured or obtained in any other way; a "time-varying waveform" is
a waveform that varies over a time dimension. Some of the
time-varying waveforms described below in relation to exemplary
implementations include intensity values, but the expression
"time-varying waveforms" also encompasses other values that vary
over time, including purely numerical values with no specified
units or other physical significance. A "sensed time-varying
waveform" is a time-varying waveform that is indicated by sensing
results obtained over time. For example, if a photosensor provides
sensed quantities that indicate intensity of received light, its
sensing results could indicate a time-varying waveform indicating
intensity sensed over time.
[0071] In a system in which sensing results, emanating light, or
other signals can include information about characteristics of
objects, an object "travels" or is caused "to travel" if the object
has a succession of positions over time with respect to one or more
parts or components of the system or one or more patterns or other
features of the system's environment such that information about
the object's traveling, e.g. about speed or other rate of
displacement, can be included in the emanating light or other
signals. An object that travels is sometimes also referred to
herein as "moving" or as having "motion" or "movement", but an
object's traveling may result from any appropriate motion of the
object and/or motion of parts or components of the system or
patterns or other features of its environment. In other words,
motion of an object includes any relative movement between the
object and parts or components of a system or patterns or features
of the system's environment, such as an encoding or sensing
component of the system or a pattern of excitation or of filtering
or another environmental pattern or feature.
[0072] A moving object's path is treated herein as providing a
directional orientation as follows: A direction parallel or
approximately parallel to the path is sometimes referred to as a
"longitudinal" or "lengthwise" direction, while a direction
perpendicular or approximately perpendicular to the path is
sometimes referred to as a "radial", "lateral", or "transverse"
direction. The lengthwise direction in which the object is moving
is sometimes referred to as "forward" or "downstream", while the
opposite direction is sometimes referred to as "backward" or
"upstream". A radial direction away from the path is "out" or
"outward", while a radial direction toward the path is "in" or
"inward". Light propagating toward the path may be referred to as
"incoming" or "incident", while light propagating away from the
path may be referred to as "outgoing". A component or arrangement
of components is "along" the path if it is disposed near the path
and has some extent in a longitudinal direction. A component or
arrangement of components is "around" the path if, in a plane
transverse to the path, it intersects multiple radial directions,
at least two of which are separated by approximately 180 degrees of
arc. A direction that similarly goes around the path is sometimes
referred to herein as a "rotation" direction. In general, it should
be understood that the above directional orientation is arbitrary
and only for ease of description, and that a moving object's path
may have any appropriate orientation.
[0073] Emanating light that includes information about an object's
traveling is sometimes referred to herein as "motion-affected"
light, as including "motion-dependent information", or as having
"motion-dependent encoding". For example, an object could travel by
being conveyed in fluid, such as liquid, gas, or aerosol, along a
path in which it emanates light that is transmitted and/or
reflected by a filter arrangement to include information about the
object's motion, thus becoming motion-affected light; in such a
case the object may be referred to as being "carried" by fluid. In
another example, an object contained in or otherwise supported by a
support structure could travel due to relative scanning movement
between the support structure and a filter component or another
component such as a photosensor, and it could emanate light that is
transmitted and/or reflected so that it becomes motion-affected
light.
[0074] The term "optical filter" or simply "filter" refers herein
to a light-transmissive or light-reflective part or component that
transmits and/or reflects light in accordance with a respective
criterion, sometimes referred to herein as a filter's "type". For
example, one general category of filters is "band pass filters",
referring to types of filters that, across some application's range
of photon energies, e.g. a range of wavelengths or frequencies such
as the visible range, preferentially transmit and/or reflect light
within a subrange, sometimes referred to as a "band"; a band pass
filter's type can therefore be specified by specifying the band or
subrange of photon energies in which it transmits and/or reflects.
A "blocking filter", which does not transmit or reflect any light
in an application's range, can be viewed as a band pass filter with
a band of zero bandwidth, while a "transparent filter", which
transmits and/or reflects all light in an application's range, can
be viewed as a band pass filter with a band that includes the
entire range.
[0075] Filters can be combined and configured in many different
ways, and all such combinations and configurations of one or more
filters are encompassed herein by the general term "filter
arrangement". A filter arrangement can include, for example, one or
more "filter components", one or more "filter assemblies", and/or
one or more "filter elements"; while the term "filter component" is
generic, referring to any component that operates as a filter, the
terms "filter assembly" and "filter element" are related and
therefore a bit more specific, in that a filter assembly is a
filter component that includes one or more filter elements, while a
filter element is a filter component that generally does not
include other filter elements within it. In general, filter
elements and filter assemblies are sometimes also referred to as
"masks". Also, the terms "transmit" and "reflect" and related
words, as used herein, include each other unless otherwise
specified, and terms such as "transmit/reflect" or
"transmitting/reflecting" encompass transmission without
reflection, reflection without transmission, and concurrent
transmission and reflection.
[0076] Filter elements of various kinds could be included in filter
assemblies, filter components, filter arrangements, and other
combinations and configurations of filters, in a wide variety of
ways. Within a given configuration of filters, relationships
between filters can be described in a number of ways. For example,
light can pass through a "sequence" of filters, meaning that
specified light passes through the filters in a sequence: If a
"radial sequence" of filters is along a path, for example,
emanating light can pass through each of the filters in the
sequence, beginning with the first and, after passing through each
preceding filter, passing through the following filter; of course,
light that is blocked by a preceding filter in a radial sequence
would not reach its following filter. If a "longitudinal sequence"
of filters is along a path, on the other hand, light emanating at
each of a sequence of segments of on the path passes through a
respective filter in the longitudinal sequence.
[0077] Several other categories of filters are described below in
relation to exemplary implementations, including shadow masks,
periodic masks, chirp masks, random masks, and so forth, and
various other categories could be identified. As used herein, the
term "random" refers to a pattern that is non-periodic over the
entire length of a longitudinal sequence of filters; in contrast, a
"periodic" filter assembly has at least one pattern that repeats
more than once across the assembly's longitudinal length; and
"chirp" patterns meet the above definition of random but can, with
linearly varying time scaling, meet the above definition of
periodic, in effect being a sequence of periods of linearly
changing frequency or wavelength. A "shadow mask" is not a band
pass filter, but rather an intensity-based filter assembly that,
within a photon energy range of interest, transmits/reflects light
of all energies, but with different parts of the filter
transmitting/reflecting the light at different intensities, such as
black and white and/or different gray scales. Any of these types of
filter assemblies can be used to obtain "spatially modulated"
emanating light, meaning emanating light that varies in time
depending on position of an object from which it is emanating.
[0078] As used herein, the term "white", in a given implementation,
refers to light with a spectrum that approximates maximum
intensities across the implementation's full range of photon
energies (which could be broad band, a combination of red, green,
and blue, or another appropriate combination); the term "black"
refers to the opposite of white, i.e. minimum available intensities
across the full range, with the ideal being no light and therefore
zero intensities. In emanating spectra, for example, light without
maximal intensities across the full range as in white may be
characterized as having a "gray level", such as one of the gray
levels between white and black, or as having a "color", such as if
it includes predominantly photon energies in a subrange, e.g. one
of the colors in the visible range of wavelengths or in the
infrared or ultraviolet ranges, or possibly a combination of such
colors. Spectra that are neither black nor white are sometimes
referred to herein as "non-binary spectra". Filter arrangement 20
includes either or both of two specified combinations or
configurations of filters: Filter component 30 includes filter
assembly 32 in which positions 34 and 36 have different
transmission functions as indicated by different cross-hatching,
while filter component 40 has a combined transmission function,
represented by box 42, in which two or more different non-uniform
transmission functions are superimposed. As a result of the
differences in transmission functions, information can be encoded
in time variation of emanating light from objects such as object
10.
[0079] As used herein, the term "transmission function" refers to a
function that indicates, for some appropriate position or set of
positions, the relationship of output and input light of a
light-transmissive and/or light-reflective component such as a
filter or filter assembly. A position's transmission function could
indicate, for example, ratio of output intensity to input intensity
at the position across a range of photon energies, sometimes
referred to herein as the transmission function's "transmission
spectrum"; a band pass filter, for example, could have
approximately the same transmission spectrum at substantially all
of its positions. A position could have any of a variety of other
kinds of transmission functions, including, for example, an
"intensity ratio", indicating the ratio of the positon's output
intensity to its input intensity, where the same intensity ratio
applies to all photon energies across the relevant range; in the
simple case in which each position of a filter has either an
intensity ratio of zero or one, each position's transmission
function could be one of a pair of binary values, such as
black/white, ON/OFF, one/zero, or the like.
[0080] Further, a band pass filter or other filter element or
assembly has a "uniform transmission function" if substantially all
its positions have transmission functions that are approximately
the same, and such a transmission function may be said to be
"approximately uniform" for light transmitted/reflected through the
filter element or assembly. Conversely, a filter element or
assembly has a "non-uniform transmission function" if its
transmission function is not approximately uniform; examples
include periodic, random, and chirp filters as described above.
[0081] Transmission functions can, of course, be different from
each other in various ways. For example, transmission functions of
two positions can differ in "color", meaning that the positions
have different transmission spectra; transmission functions with
transmission spectra that have the same shape across a relevant
range can differ in "intensity", meaning that they have different
intensity ratios. Similar terminology can be applied to uniform
transmission functions for filter elements, components, or
assemblies. In FIG. 1, elements 34 and 36 have transmission
functions that are different from each other. Also, simpler
transmission functions that are superimposed to provide a combined
transmission function can have different transmission functions; in
FIG. 1, two of the simpler transmission functions superimposed to
provide combined transmission function 42 are non-uniform and
different from each other.
[0082] Different transmission functions can also be combined in a
number of ways. For example, in a longitudinal sequence of filters,
transmission functions are similarly combined into a sequence. In a
stack or other radial sequence of filters or filter assemblies, on
the other hand, transmission functions can be "superimposed",
meaning that both transmission functions are applied to light
passing through the component, resulting in a combined transmission
function in which simpler transmission functions are superimposed.
As used herein, a transmission function is "simpler" than a
combined transmission function in which it is superimposed with at
least one other transmission function, except in cases where the
combined transmission function and all of the superimposed
transmission functions have the same spectral shape or where the
superimposed transmission functions have related shapes that result
in uniform loss of detail when superimposed in specific phase
relationships; although there are many abstract examples of
superpositions that result in uniform loss of detail (e.g. two
square waves of the same period and at opposite phase would have a
flat line superposition) simplifying superpositions are very
unlikely to occur between transmission functions with disparate
shapes, such as random and periodic, random and chirped, chirped
and periodic, and so forth--some detail might be lost locally in
such cases, but most detail is preserved. Simpler transmission
functions can be superimposed to obtain a combined transmission
function in various ways other than a stack or radial sequence; for
example, as described below in relation to some exemplary
implementations, a single filter assembly can have a combined
transmission function that is "stack-equivalent", meaning that it
is approximately equivalent to a stack of filter components with
simpler transmission functions. In some cases, including certain
types of reflective filters, a stack-equivalent filter assembly can
be equivalent to a combination of simpler filters without regard to
the order in which they are superimposed, so that it is equivalent
to a number of different stacks in which the simpler filters are in
different orders.
[0083] As shown within combination 30, when object 10 is in segment
50 or segment 52 of its path, respective portions of emanating
light are transmitted/reflected through positions 34 and 36 of
filter assembly 32, as indicated respectively by arrows 54 and 56.
Because of the different transmission functions, this
transmitting/reflecting operation encodes information in time
variation of the emanating light, represented by arrows 58.
Specifically, if the emanating light from segments 50 and 52 has
the same intensity or spectrum, the output light from positions 34
and 36 can affect its intensity or spectrum differently, and this
difference can indicate, for example, the time at which object 10
moved between segment 50 and segment 52. In the illustrated
example, segment 50 precedes segment 52 along the path of object
10, so that the portion of emanating light from segment 50 is
transmitted/reflected according to the transmission function of
position 34 before the portion of emanating light from segment 52
is transmitted/reflected according to the transmission function of
position 36.
[0084] In some exemplary implementations below, for example, a
filter assembly can have a longitudinal sequence of band pass
filter elements with bands of different colors. As a result, output
light from filter elements of different colors will have different
intensities, depending on the spectrum of light emanating from an
object, so that time variation of the output light encodes
information about the emanating light's spectrum, i.e. about the
type of the object. In other examples, information about speed or
other displacement rate and position can be encoded by longitudinal
filter sequences.
[0085] As shown within component 40, on the other hand, as object
10 passes through each of a series of segments that includes
segments 60 and 62, respective portions of emanating light are
transmitted/reflected with combined transmission function 42, as
indicated by respective arrows 64 and 66. Because at least two of
the simpler transmission functions that are superimposed in
function 42 are non-uniform and different from each other, this
transmitting/reflecting operation also encodes information in time
variation of the emanating light, represented by arrows 68. If the
emanating light from each segment in the series has the same
intensity or spectrum, the output light from function 42 will be
encoded in accordance with both of the simpler non-uniform
transmission functions. In other words, information in accordance
with both of the simpler transmission functions will be
concurrently encoded in time variation of the emanating light.
[0086] In some exemplary implementations below, for example, a
stack or stack-equivalent filter assembly combines a periodic or
chirp transmission function that can encode information about an
object's position, speed, or other displacement rate with a random
transmission function that can encode information about an object's
spectrum or type. Emanating light passing through the filter
assembly is concurrently encoded with both types of
information.
[0087] As suggested by the words "AND/OR" between combination 30
and component 40, the two are not mutually exclusive, and could be
implemented together. As described below in relation to some
exemplary implementations, a single filter assembly could encode
information in time variation of emanating light in both of the
ways illustrated for combination 30 and component 40.
[0088] The operation in box 70 photosenses the emanating light that
has information encoded in its time variation, represented by
arrows 58 and 68. This operation can be implemented with any
suitable photosensing component, some of which are described below.
In general, sensing results from photosensing take the form of
analog or digital electrical signals, depending on the structure
and circuitry included in the photosensing component. The operation
in box 72 uses the sensing results from box 70 to obtain data
indicating some or all of the encoded information, and can
therefore be referred to as a "decoding" operation. The results of
decoding can be used in a wide variety of ways, some of which are
described below in relation to specific implementations.
[0089] Information about an object, as obtained in FIG. 1, can be
used for a wide variety of purposes. In exemplary implementations
described below, such information can, for example, be used to
distinguish objects. In some applications, such as where the
distinguished objects are registration marks in documents or other
images, appropriate subsequent operations can be controlled based
on the results of distinguishing objects.
[0090] Filtering arrangement 20 in FIG. 1 could be implemented in
many different ways, some of which are described below. In some
exemplary implementations below, for example, a filter component
includes positions that have different transmission functions. In
others, a filter assembly has a combined transmission function with
superimposed simpler, non-uniform transmission functions. These
techniques can be implemented together. As a result of these
techniques, emanating light will have time variation due to
different transmission functions, and the time variation of the
emanating light can encode information about the object's spectral
interactions such as the spectra in which it and other similar
objects absorb, fluoresce, or otherwise interact with light, i.e.
about the type of the object.
[0091] FIG. 2 schematically illustrates general features of system
100, a system in which light emanating from a moving object can
include information about characteristics of the object and in
which features described above in relation to FIG. 1 can be
implemented. As with other exemplary implementations described
below, system 100 involves a combination of parts or components.
Encoding component 102 illustratively provides output light that
includes information about one or more object characteristics.
Photosensing component 104 responds to the output light, providing
sensing results such as electrical output signals with information
in a form that can be communicated to processing component 106,
possibly after conversion to other forms, e.g. for storage,
transmission, and processing, such as optical or other
electromagnetic signal forms. Processing component 106 can use the
sensing results from photosensing component 104 to obtain and/or
provide characteristic data indicating information about one or
more object characteristics.
[0092] Object 110 illustratively travels in a direction indicated
by arrows 112, passing through a succession of positions, two of
which are illustrated. In some positions, object 110 can receive
excitation, illustrated by arrows 114, and, in response, light as
illustrated by arrows 116 can emanate, such as from fluorescence of
a dye or other "tag" attached to object 110 or from native
fluorescence or autofluorescence of object 110 itself, e.g. due to
ultraviolet light or other excitation of intrinsic cell material or
other material in object 110; except as otherwise noted, however,
implementations described herein can additionally or alternatively
employ chemofluorescence, biofluorescence, absorption, scattering,
or other phenomena that do not require concurrent excitation. More
generally, excitation could take any appropriate form and is not
limited to illumination, and excitation and emanation need not be
concurrent or otherwise coincident, but could have any appropriate
relationship in space and time. Some examples of excitation are
described below in relation to exemplary implementations.
[0093] Arrow 120 represents output light from encoding component
102. Box 122 between arrows 116 and arrow 120 illustrates that
information about one or more characteristics of object 110 is
included in the output light. As described below in relation to
exemplary implementations, this information can be encoded in a
variety of ways, including, for example, patterning excitation
and/or patterning emanating light to obtain encoded output light
represented by arrow 120.
[0094] Arrow 120 points to photosensing component 104, indicating
that at least part of the encoded output light is illustratively
sensed by component 104 to obtain sensing results. Based on the
sensing results, component 104 provides electrical output signals
represented by arrow 130. The electrical output signals can also
include at least some of the information about object
characteristics from box 120. As a result, processing component 106
can, in response to the electrical output signals, obtain and/or
provide characteristic data indicating information about object
characteristics.
[0095] Each of components 102, 104, and 106 in FIG. 2 could be
implemented in a wide variety of different ways. FIGS. 3-5
illustrate several general features of implementations of encoding
component 102, each of which involves an arrangement along a path
traveled by a moving object.
[0096] In FIG. 3, excitation arrangement 150 is along a path
traveled by moving object 152 as it emanates light within an
encoding component such as component 102 in FIG. 2. As suggested by
the one-dimensional coordinate axis labeled "x OR t", the path can
be treated either as extending in space, such as along an
x-direction, or as occurring over time, t; unless otherwise
indicated hereafter in relation to a specific exemplary
implementation, the x-direction refers to an object's path and
therefore might not in some cases follow a straight line relative
to the environment. Although the speed or other rate of
displacement of object 152 may vary as it travels along the path,
information about its speed or other rate of displacement can be
sufficient to allow an approximate mapping between its x-direction
positions and times t; more generally, mapping between an object's
x-direction positions and times t can be based on any suitable
system, such as with trigger detection techniques as described in
U.S. Patent Application Publication No. 2007/0145249, entitled
"Sensing Photons from Objects in Channels", incorporated herein by
reference in its entirety, or from other techniques, including
obtaining information such as a trigger signal from an object's
encoded signal.
[0097] Although excitation components could be positioned in any
appropriate way along a path, the exemplary implementations
described below generally involve arrangements of one or more
excitation components along the x OR t axis, and FIG. 3 shows
several exemplary components within a sequence of K excitation
components 154 through 156, with component 154 labeled "0" and
component 156 labeled "(K-1)". Excitation components need not,
however, be arranged on only one side of the path, but rather could
be positioned at any suitable positions around the path, depending
on how excitations from different components interact. Also, two or
more excitation components could be at the same position or in
overlapping position ranges along the x OR t axis, but displaced in
a rotation direction; a configuration of excitation components that
are sufficiently displaced in a rotation direction so that they are
around the path is illustrated by component 158, representing a
possible position of another excitation component labeled "(0')" in
arrangement 150, on the opposite side of the path traveled by
object 152 from component 154.
[0098] Arrow 160 schematically represents excitation from component
154, while arrow 162 represents excitation from component 158.
Similarly, arrow 164 represents excitation from component 156.
Although excitation from components 154 and 158 can be provided
concurrently to object 152, as suggested by arrows 160 and 162,
excitation from component 156, represented by arrow 164, is
provided at a subsequent position and time of object 152.
[0099] Excitation component 170, labeled "k1", illustratively
includes one or more interfering light sources 171, resulting in
two or more different types of excitation, with two types
represented by arrows 172 and 174. The excitation represented by
arrow 172 occurs while object 152 travels along a segment of the
path through region 176, while the type of excitation represented
by arrow 174 occurs while object 152 travels along a subsequent
segment of the path through region 178. Regions 176 and 178
therefore form a pattern in space, an example of "spatially
patterned excitation" used herein to refer to excitation that
occurs in a pattern in space, i.e. a "spatial pattern"; spatially
patterned excitation could, for example, include multiple periods
of a spatial pattern. In particular, the excitation in region 176
has a different photon energy spectrum than the excitation in
region 178, so that regions 176 and 178 could be described as
having "different colors" of excitation. Several specific examples
in which spatially patterned excitation includes regions of
different colors are described below in relation to exemplary
implementations; as will be understood from some of the examples,
the x-direction of a path as shown in FIG. 3 may not follow a
straight line, so that regions 176 and 178 may not in fact be
oriented along a straight line through components 154 through
156--in some implementations, regions 176 and 178 could each extend
parallel to such a line and the path could go back and forth
between regions 176 and 178.
[0100] Excitation component 180, labeled "k2", illustratively
includes one or more structured light sources 182. In other words,
light sources 182 are structured to provide spatially patterned
excitation, represented by spatial pattern 186. In the illustrated
example, arrow 184 represents excitation provided in region 188,
one of a pattern of regions through which object 152 passes while
receiving excitation from component 180. The complete pattern of
regions is represented in FIG. 3 by pattern 186.
[0101] FIG. 3 also illustrates lines 190 through which each of
components 154 through 156 can receive control signals from
excitation control circuitry (not shown). For example, one or more
of the components in excitation arrangement 150 could include
trigger detecting circuitry (not shown) as described above, and the
excitation control circuitry could, in response to the trigger
detecting circuitry, provide control signals causing the component
to provide excitation, either in a steady state or time-varying
manner. As described below in relation to exemplary
implementations, time-varying excitation can encode information in
a way similar to spatially patterned excitation.
[0102] Additional description of excitation techniques is set forth
in co-pending U.S. patent application Ser. No. 12/023,436, entitled
"Producing Time Variation in Emanating Light", incorporated herein
by reference in its entirety.
[0103] In FIG. 4, filter arrangement 200 is similarly along a path
traveled by moving object 202 as it emanates light within an
encoding component such as component 102 in FIG. 2. Filter
arrangement 200 includes a combination of one or more filter
assemblies along the path traveled by object 202.
[0104] Although filter assemblies could be positioned in any
appropriate way along a path, the exemplary implementations
described below generally involve arrangements of filter assemblies
along the x OR t axis, and FIG. 4 shows several exemplary cross
sections of filters within a sequence of M filter assemblies 204
through 206, with each cross section being taken parallel to the x
OR t axis and with assembly 204 labeled "0" and assembly 206
labeled "(M-1)". Filter assemblies need not, however, be arranged
on only one side of the path as shown, but rather could be
positioned at any suitable positions around the path, depending on
directional intensity variations of emanating light. Also, two or
more filter assemblies could be at the same position or in
overlapping position ranges along the x OR t axis, but displaced in
a rotation direction; a configuration of filter assemblies that are
sufficiently displaced in a rotation direction so that they are
around the path is suggested by box dashed-line box 208 in FIG. 4,
representing a possible position of another filter assembly labeled
"(0')" in arrangement 200, on the opposite side of the path
traveled by object 202 from filter assembly 204.
[0105] Filter assembly 210, labeled "m1", illustratively includes a
radial sequence of filters through which light emanating from
object 202, represented by arrow 212, can pass, with the output
light from filter assembly 210 being represented by arrow 214.
Filter assembly 210 could include any appropriate number of
filters, with filters 216 and 218 being shown in FIG. 4.
[0106] The overall sequence of filter assemblies 204 through 206
illustrates a longitudinal sequence. Further, filter assembly 220
includes a longitudinal sequence of filters through which light
emanating from object 202, represented by arrows 222, can pass,
with the output light from filter assembly 220 being represented by
arrows 224. Filter assembly 220 could include any appropriate
number of filters in any appropriate longitudinal sequence, with
adjacent filters 226 and 228 being shown in FIG. 4. Each of filters
226 and 228 could, for example, be a band pass filter, with the
bands of filters 226 and 228 being sufficiently different to
provide useful information about an emanation spectrum of object
202. Such a filter assembly is sometimes referred to herein as a
"spatially patterned filter", because the filters it includes can
be treated collectively as a single filter that has a pattern that
varies as a function of position. Several examples of spatially
patterned filters are described below in relation to exemplary
implementations, and one or both of filters 216 and 218 in assembly
210 could also be implemented as a spatially patterned filter.
[0107] In the specific example of filter assembly 220, output light
per arrows 224 can include encoded information from filters 226 and
228, and the encoded information can be recovered by photosensing
the output light and performing appropriate operations on the
sensing results. In general, filters 226 and 228 and other filters
in filter assembly 220 can have any suitable lengths in the x OR t
direction that allow recovery of the encoded information by
photosensing and signal processing, including lengths smaller than
the apparent extent of object 202 in the x OR t direction that may
result in some loss of resolution analogous to blurriness or
smearing. As described in relation to some exemplary
implementations below, however, each of filters 226 and 228 can
have length in the x OR t direction greater than or equal to an
apparent extent of object 202 in the x OR t direction, while the
lengths of filters 226 and 228 (and other filters in assembly 220)
can be sufficiently small that characteristics of object 202
indicated by emanating light do not change while object 202 is
traveling past assembly 220. In some specific implementations,
filters 226 and 228 have parallel sides extending in a direction
transverse to the path, and an assembly of such filters is
sometimes referred to herein as a "striped filter" in which each
stripe can be specified by filter type and its length (or width) in
the lengthwise direction.
[0108] In the specific example of filter assembly 220, output light
per arrows 224 can include encoded information from filters 226 and
228, and the encoded information can be recovered by photosensing
the output light and performing appropriate operations on the
sensing results. In general, filters 226 and 228 and other filters
in filter assembly 220 can have any suitable lengths in the x OR t
direction that allow recovery of the encoded information by
photosensing and signal processing, including lengths smaller than
the apparent extent of object 202 in the x OR t direction that may
result in some loss of resolution analogous to blurriness or
smearing. As described in relation to some exemplary
implementations below, however, each of filters 226 and 228 can
have length in the x OR t direction greater than or equal to an
apparent extent of object 202 in the x OR t direction, while the
lengths of filters 226 and 228 (and other filters in assembly 220)
can be sufficiently small that characteristics of object 202
indicated by emanating light do not change while object 202 is
traveling past assembly 220. In some specific implementations,
filters 226 and 228 have parallel sides extending in a direction
transverse to the path, and an assembly of such filters is
sometimes referred to herein as a "striped filter" in which each
stripe can be specified by filter type and its length (or width) in
the lengthwise direction.
[0109] Filter arrangements similar to those shown in FIG. 4 may
find application not only in fluidic implementations as described
below but also in implementations in which objects in an array move
relative to other components due, for example, to scanning
movement. One such area of application is in image scanning, such
as with scanning sheets of paper or other media that can bear
images. In particular, object 202 could be a colored spot on a
sheet of paper or other medium, and a filter arrangement could be
used to obtain information about small differences in color of
light emanating from object 202, e.g. color of reflected light in
response to broadband illumination. Such information could be used
to obtain position and/or color of object 202; for example, if
object 202 is a registration mark with a color unique to
registration marks, its color could be accurately distinguished
from spots of other colors using techniques as described herein and
its position could be obtained with sufficient accuracy to allow
registration of the sheet, whether for image sensing or for
printing or another operation on the sheet. Very high accuracy
sensing of color is sometimes referred to as "hyperspectral color
sensing".
[0110] In FIG. 5, displacement control arrangement 250 is similarly
along a path traveled by moving object 252 as it emanates light
within an encoding component such as component 102 in FIG. 2.
Displacement control arrangement 250 includes a combination of one
or more displacement control components, each of which is
illustratively shown enclosing a respective segment of the path
traveled by object 252. It would, of course, be possible to
implement display control components in other ways, such as where
an object travels along a path that is not enclosed within a
channel or fluidic structure.
[0111] Although displacement control components could be positioned
in any appropriate way along a path, the exemplary implementations
described below generally involve arrangements of displacement
control components along the x OR t axis, and FIG. 5 shows several
exemplary components within a sequence of control components 254
through 256, with component 254 labeled "0" and component 256
labeled "(N-1)". Although each displacement control component in
the sequence illustratively contains a respective segment of the
path, it may be possible to implement displacement control
components that affect displacement in overlapping segments of a
path or that interact in other ways.
[0112] Control component 260, labeled "n1", illustratively includes
shaped boundary 262, meaning that a boundary that extends partially
or completely around the path, such as the boundary of a fluidic
channel, has a shape that affects or controls displacement of
object 252 as it travels along the path, such as by affecting its
speed or other rate of displacement. Several examples of boundary
shapes are described below in relation to exemplary
implementations.
[0113] Control component 270, labeled "n2", illustratively includes
motion device 272. Device 272 can illustratively cause lateral
motion of a boundary in its segment of the path, as suggested by
bidirectional arrows 274. Line 276 shows that device 272 can
receive control signals from displacement control circuitry (not
shown). Component 270 could also include trigger detecting
circuitry (not shown), and the displacement control circuitry could
respond to the trigger detecting circuitry by initiating operation
of device 272, either in a steady state or time-varying manner.
Examples of how device 272 could be implemented are described below
in relation to specific implementations.
[0114] FIG. 6 illustrates system 400, an exemplary system that
could implement components as in system 100 in FIG. 2. Although
system 400 illustratively includes central processing unit (CPU)
402 connected to various components through bus 404, a wide variety
of other architectures could be employed, including any appropriate
combination of hardware and software, as well as specialized
hardware components such as application specific integrated
circuits (ASICs) for one or more of the illustrated components or
in place of a software component executed by CPU 402. Furthermore,
CPU 402 could be the CPU component of any suitable machine such as
a laptop or desktop computer, a specialized computer for system
400, and CPU 402 and other digital components as shown could be
replaced by other specialized circuitry, such as an analog signal
processor; in a relatively simple application, CPU 402 could be
implemented with a single digital signal processor or a CPU of a
laptop or other personal computer receiving time-varying signals.
On the other hand, in some applications, it may prove advantageous
to implement all signal processing with analog circuitry, including
operations that compare time-varying waveforms and that obtain
their derivatives or other related waveforms, making it possible to
replace substantially all the digital components as shown if
appropriate.
[0115] System 400 also includes external input/output (I/O)
component 406 and memory 408, both connected to bus 404. External
I/O 406 permits CPU 402 to communicate with devices outside of
system 400.
[0116] Additional components connected to bus 404 are within or
connected to system 400. In the illustrated implementation of
system 400, IC I/O 410 is a component that permits CPU 402 to
communicate with ICs such as photosensing ICs; M ICs are
illustrated in FIG. 6 by a series extending from IC(0) 412 to IC
(P-1) 414. ICs 412 through 414 illustratively include IC(p) 416
with a photosensor array 418, which includes photosensing cells.
Similarly, device I/O 420 is a component permitting CPU 402 to
communicate with various devices in system 400, such as sensing and
control devices; Q devices in system 400 are represented in FIG. 6
by device (0) 422 through device (Q-1) 424. In addition to
excitation components as described above in relation to FIG. 3 and
displacement control components as described above in relation to
FIG. 5, devices 422 through 424 can include fluidic devices such as
pumps, metering electrodes, smart gates, and other devices for
gating and bifurcating, valves, flow or pressure sensors, and so
forth. Such fluidic devices could be implemented in various ways;
smart gates, for example, could be implemented with MEMS style
microgates or by using electromagnetic forces, which are effective
because most particles are charged such that an electric field can
be used to direct them as desired in a channel.
[0117] Memory 408 illustratively includes program memory 430
although instructions for execution by CPU 402 could be provided in
various other forms of software or hardware, on or off of CPU 402.
The routines stored in program memory 430 illustratively include
encoding routine 440; detect, readout, and combine routine 442; and
object distinguishing routine 444. In addition, program memory 430
can also store a number of subroutines (not shown) that CPU 402 can
call in executing routines 440, 442, and 444.
[0118] CPU 402 executes encoding routine 440 to encode information
in light emanating from a moving object as it travels a path, i.e.
information about characteristics of the object. In doing so,
routine 440 can provide receive input signals from and provide
output signals to devices 422 through 424. For example, to obtain
appropriate motion of the object, CPU 402 can receive signals from
sensors, perform computations to determine what fluidic operations
are necessary, and then provide signals to activate pumps, metering
electrodes, gates, and valves to produce appropriate relative
movement between an object and other components of system 400 along
its path. CPU 402 can also receive signals from trigger detecting
devices and perform computations to determine what control signals
to provide to excitation components, motion devices, or other
components or devices in order to perform appropriate encoding in
emanating light. Several examples of techniques that can be
performed by encoding routine 400 are described below in relation
to exemplary implementations.
[0119] In executing routine 442, CPU 402 can, for example, perform
pre-sensing readout, obtain object information and sensing periods,
perform sensing readout with sensing periods and analog adjustment,
digitally adjust sensing results and store quantities for an
object, and combine the quantities for an object to produce its
characteristic data. Routine 442 could, for example, call a
subroutine implemented as described in U.S. Patent Application
Publication Nos. 2007/0145249, entitled "Sensing Photons from
Objects in Channels", and 2007/0146704, entitled "Sensing Photons
Energies Emanating from Channels or Moving Objects", each of which
is incorporated herein by reference in its entirety. Such a
subroutine can be implemented for single objects moving past arrays
or for spaced multiple objects moving past arrays, provided
spacings between objects are sufficient to avoid interference.
Also, such a subroutine can follow a general strategy of performing
a series of readout operations, after which information for an
object is combined and its characteristic data is provided,
although it would also be possible to provide the information from
each readout operation immediately.
[0120] FIG. 7 illustrates an example of how object distinguishing
routine 444 in FIG. 6 could be implemented, using each object's raw
data from routine 442 before it is used to obtain characteristic
data for the object. Routine 444 can begin with the operation in
box 470, which coordinates routines 440 and 442 as described above,
obtaining an object's raw data, such as a data structure with
photosensed quantities obtained from ICs 412 through 414.
[0121] The operation in box 472 receives the raw data from box 470,
such as in the form of a handle or other item of data necessary to
access a data structure. The operation in box 472 then uses the raw
data to obtain the object's characteristic data, such as in one of
the ways described below in relation to exemplary implementations.
For example, an appropriate comparison technique could be used to
obtain a comparison result indicating an object's type or other
characteristic. The characteristic data from box 472 can indicate
whether the object is of interest for further analysis, such as
because it may be suspicious or harmful or, on the other hand,
because it may be of interest for more refined analysis.
[0122] The operation in box 480 branches based on whether the
object is of interest. If not, the operation in box 482 opens a
smart gate or provides appropriate control signals to perform
another operation to purge the object from the system. But if the
object is of interest, the operation in box 484 ensures that the
smart gate is closed or provides control signals for other suitable
operations to transfer the object downstream so that a more refined
or detailed analysis or other further analysis can be performed,
possibly after concentration of the object with other similar
objects by appropriate fluidic devices.
[0123] FIG. 8 illustrates an application of a system as in FIGS. 6
and 7 in analyzer 500 on support structure 502, a fluidic
structure. Defined in support structure 502 is serpentine channel
504 through which object 506 can travel, carried by fluid such as
liquid, gas, or aerosol or moved in some other appropriate way.
Object 506 can, for example, be a biological cell or another object
of any of the types mentioned above.
[0124] The manner in which object 506 enters channel 504 and is
carried by fluid can be the same as described in U.S. Patent
Application Publication Nos. 2007/0145249, entitled "Sensing
Photons from Objects in Channels", and 2007/0146704, entitled
"Sensing Photons Energies Emanating from Channels or Moving
Objects", each of which is incorporated herein by reference in its
entirety. As explained there, object 506 can be carried through
channel 504 by operation of propulsion components and can be purged
or otherwise caused to exit, together with fluid that is carrying
it, from one of several outlets, such as through toggling of
valves. While in channel 504, object 506 can travel through a
series of sensing components, each of which can obtain information
about object 506.
[0125] The first two sensing components after object 506 enters
channel 504 are illustratively Coulter counter 510, an electrically
based particle size detector, and Mie scatter sensor 512, also a
particle size detector. Information about size of object 506 from
Coulter counter 510 and Mie scatter sensor 512 can be used in
obtaining information about its other characteristics.
[0126] The next sensing component along channel 504 is emanating
light encoder/photosensor 520, shown schematically in a
cross-sectional view along an axis similar to the x OR t axis in
FIGS. 3-5, although it would typically be implemented instead with
components above and below channel 504, similarly to other sensing
components described below. The schematic illustration of
encoder/photosensor 520 includes excitation/displacement component
522, filter component 524, and photosensing component 526, all of
which might be implemented in a variety of ways, including some of
those described above and below.
[0127] After passing through encoder/photosensor 520, object 506
could be characterized without obtaining further information, or,
as in the illustrated implementation, object 506 can continue
through subsequent sensing components, illustratively including
components 530, 532, and 534. These could, for example, include
first and second fluorescence sensing components and a Raman
scatter sensing component. Information obtained from any
combination of the sensing components can be used to distinguish
between types of objects, such as different types of biological
cells, or to distinguish objects from environment or background.
Based on such a distinction, valve 540 at a bifurcation junction
can be toggled between two positions, with object 506 exiting as
indicating by arrow 542 if valve 540 is in one position and exiting
as indicated by arrow 544 if valve 540 is in another position.
[0128] The fluidic implementation in FIG. 8 is merely illustrative
of a wide variety of implementations of the techniques described
herein. For example, any appropriate fluidic or nonfluidic
techniques could be used with a wide variety of different types of
objects and various types of relative motion to gather various
types of information about object characteristics.
[0129] FIG. 9 illustrates an example of article 600 with components
that could be operated similarly to encoder/photosensor 520 in FIG.
8. Some features of article 600 can be understood from description
in co-pending U.S. patent application Ser. No. 11/777,712, entitled
"Producing Fluidic Waveguides", incorporated herein by reference in
its entirety. For example, article 600 includes a "fluidic
structure", used herein to refer to a structure that depends for
its operation on fluid positioning or fluid flow, such as, for
liquids or gases, in response to pressure or, for liquids, as a
result of surface tension effects; in general, the term "fluid" is
used herein to encompass all media that can flow, including
liquids, gases, aerosols, and so forth. The related term "channel"
refers herein to any tube or other enclosed passage within a
fluidic structure through which fluid flows during operation. A
channel is therefore an example of a "fluidic region", used herein
to refer to a region that can contain fluid. An operation
"positions" fluid in a channel if it changes the fluid's position
in any way that leaves the fluid in the channel.
[0130] A channel or portion of a channel through which objects can
travel along paths are treated herein as having the directional
orientation described above in relation to a path. In addition, a
"cross section" lies in a plane perpendicular to a direction in
which a local net flow of fluid through the channel or portion can
occur; a direction in which a cross section extends can be referred
to as a "transverse direction" or a "lateral direction." A channel
or portion with approximately uniform cross section and
substantially linear longitudinal direction can be referred to as
"straight", and the channels and portions described herein are
generally straight unless otherwise indicated.
[0131] In order to contain fluid, a channel or other fluidic region
is typically "bounded", meaning that surfaces or surface areas
bound it on at least some sides. A "boundary" of a channel or
portion is the surface or combination of surfaces within which
fluid contained in the channel is confined. A "port" is an opening
that extends through the boundary of a channel or portion such that
fluid can enter or exit through the port; in general, a port is
relatively small compared to the length of the channel or portion,
and the boundary is treated as extending across the port as if the
port did not exist.
[0132] As described below, article 600 can include two
light-transmissive components, and FIG. 9 shows article 600 in a
top view through one light-transmissive component. In this view,
the inner region between the light-transmissive components includes
two main portions, channel portion 602 that can contain fluid and
non-channel portion 604 that surrounds channel portion 602; channel
portion 602 is illustratively shaped like a "T", but could instead
have an L-shape or any other suitable shape, including a serpentine
shape as in FIG. 8. Ports 608 are openings through one of the
light-transmissive components, allowing entry and exit of fluid
into and out of channel portion 602.
[0133] FIG. 9 also shows filter assembly 610 in dashed outline.
Filter assembly 610 is illustratively a spatially patterned filter
with a longitudinal sequence of band pass filters that includes
filters 612, 614, 616, 618, and 620. Filters 612, 616, and 620 are
illustratively cross-hatched similarly to each other to indicate
that they have the same or approximately the same band, while
filters 614 and 618 are also cross-hatched similarly to each other,
illustrating that they also have the same or approximately the same
band, a band that is different than that of filters 612, 616, and
620. In other words, filter assembly 610 is a striped filter in
which each of filters 612 through 620 can be specified by the band
that it passes and its length in the x-direction in FIG. 9.
[0134] Surrounding filter assembly 610, blocking material 622 is
structured and positioned to provide an aperture. Blocking material
622 can, for example, be a material with approximately zero light
transmission that prevents scattering and reflection of light, also
preventing light entering filter assembly 610 from nearby
fluorescing objects. Blocking material 622 can be produced during
the same operation that produces filters 612 through 620 and can in
effect be part of filter assembly 610.
[0135] The cross section in FIG. 10 shows how light-transmissive
components 630 and 632 are separated by material in non-channel
portion 604. For example, components 630 and 632 can each include
quartz or another suitable material such as glass or acrylic with
an appropriate thickness; in a successful implementation, for
example, component 630 has a thickness of approximately 0.3 mm,
while component 632 has a thickness of approximately 1.0 mm or
less; depending on the application, on stability of materials used,
and size of objects being characterized, suitable thicknesses might
range from a few millimeters down to 0.1 mm or even less. The
optimum distance between them is determined primarily by the size
of objects being characterized. For biological cells with typical
dimensions of 10 .mu.m, for example, the distance can be
approximately 20 to 50 .mu.m, maintained by material in non-channel
portion 604, which could, for example, be a suitable photoresist
material such as SU-8 or another polymer material. Alternatively, a
wall (not shown) could be formed around channel portion 602, and
non-channel portion 604 could then be filled with epoxy material
that seals a lateral boundary around channel portion 602. Various
other techniques could be used to produce a similar fluidic
structure, including hot embossing, nano-imprinting, or injection
molding, and channel portion 602 can have appropriate dimensions,
such as for waveguiding as described in co-pending U.S. patent
application Ser. No. 11/777,712, entitled "Producing Fluidic
Waveguides", incorporated herein by reference in its entirety.
[0136] FIG. 10 also shows object 640 from which light is
illustratively emanating upward, as illustrated by an emission
cone. Although the emission cone is illustratively shown as a
single cone, the actual emission cone would depend on angles of
total internal reflection at surfaces through which emanating light
is transmitted in article 600. FIG. 10 illustrates three
alternative filter assembly positions, with filter assembly 642
facing channel portion 602, on the lower surface of component 630;
with filter assembly 644 being outside of channel 602 on the upper
surface of component 630; and with filter assembly 646 being spaced
apart from the upper surface of component 630, adjacent photosensor
648, which could, as in other implementations, be a single, large
area photosensor (such as a photo-diode, an avalanche photo-diode
(APD), or a photo-multiplier tube (PMT)), or an appropriate array
of photosensing cells whose sensed quantities can be combined to
obtain a single photosensed quantity, such as an intensity value
for a sensing period. As suggested in FIG. 10, the emission cone
from object 640 is imaged onto image plane 650 extending through
filter assembly 646 by optical component 652, illustratively shown
as a single lens, but which could be any suitable lens, lens
system, or other optical component, some examples of which are
described in co-pending U.S. patent application Ser. No.
11/698,409, entitled "Method and System Implementing Spatially
Modulated Excitation or Emission for Particle Characterization with
Enhanced Sensitivity", incorporated herein by reference in its
entirety.
[0137] The emission cone for filter assembly 642 includes the range
of angles of incident light that are not totally reflected by the
surface of assembly 64. Similarly, the emission cone of filter
assembly 644 is determined by the range of angles within which
emanating light is not subject to total internal reflection at the
surface between component 630 and assembly 644. The emission cone
for filter assembly 646 is similar to that for filter assembly 644,
but can occupy a smaller area on filter assembly 646 due to the
effect of optical element 652.
[0138] In one illustrative example, channel portion 602 contains
water with an index of refraction n=1.33, and object 640 has a
diameter d=7 .mu.m, which would be typical for certain biological
cells, e.g. T-lymphocytes. Channel portion 602 has a height between
components 630 and 632 of 30 .mu.m and its distance from the lower
surface of filter assembly 642 is approximately h=15 .mu.m.
Component 630 is acrylic with an index of refraction n=1.48,
surrounded by air with an index of refraction n=1. If filter
assembly 642 were absent, the escape angle from channel portion 602
to component 630 would be .alpha.(escape)=48.75.degree., which
would determine the size of the emission cone in which light from
object 640 can leave channel portion 602. The angle of total
internal reflection at the upper surface of component 630, on the
other hand, can be obtained as .alpha.(TIR)=42.51.degree., which
determines the size of the emission cone for light that leaves
component 630. The diameter of a disk illuminated by object 640 at
the water-acrylic interface can be obtained from
D=d+2*h*tan(.alpha.(escape))=(7+(2*17.1)).mu.m=41.2 .mu.m, where
17.1 .mu.m is the radius of the maximum emission cone that can pass
through component 630 without total internal reflection. The
"minimum feature size" ("MFS") for a pattern suitable to detect
object 640 at the water-acrylic interface would be equal to D or
approximately 40 .mu.m; in general, MFS can be defined for a mask
along the path of an emanating particle as the extent in the path's
longitudinal direction of the mask's smallest uniform feature (i.e.
the smallest transmitting filter element or the smallest blocking
filter element, whichever is smaller).
[0139] Where photosensor 648 is implemented with a numerical
aperture that makes the emission cone smaller, filter assembly 642
can accordingly have a slightly smaller MFS than calculated as
above; similarly, in some acrylic implementations of component 630,
some light typically leaves component 630 at an angle slightly
higher than .alpha.(TIR), which could also allow a slightly smaller
MFS. In general, however, the MFS of filter assembly 642, if too
small, results in passage of light from an object's emission cone
around both sides of a feature in assembly 642, so that the
time-varying signal of a photosensor, while containing some
information, may not accurately indicate information about
displacement of the object as it travels along a path past filter
assembly 642. Similar considerations apply to filter assemblies 644
and 646, with the MFS of filter assembly 644 necessarily being
significantly larger than that of filter assembly 642, but with the
MFS of filter assembly 646 possibly being intermediate between
those of assemblies 642 and 644, depending on the precision of
optical component 652. In implementations without optical
components, photosensor 648 could be slightly larger due to
spreading of emanating light. For a biological cell on the order of
10 .mu.m, a typical MFS would be in the range of 10-20 .mu.m. The
channel width might be an order of magnitude larger, while the
channel length might be two orders of magnitude larger, and the
width of the filter assembly would depend on the channel width. For
example, assembly 642 might be 100 .mu.m wide and approximately 1.0
mm long. At the time of manufacture, a calibration operation could
be performed using objects that are, for example, tiny beads with
known fluorescence spectra; light emanating from such beads could
be measured and used to obtain calibration values necessary to
adjust measured values to obtain known intensities for such
objects.
[0140] The cross section in FIG. 11 further illustrates how
component 630 has oblique surface 660, a light interface surface
that is illustratively at an angle of approximately 45.degree. to
the inward-facing surfaces of components 630 and 632. As a result,
incident excitation light at a direction approximately
perpendicular to surface 660, as illustrated by arrow 662, can
cause and couple with light propagating through channel portion
602, as illustrated by arrow 664, as described, for example, in
co-pending U.S. application Ser. No. 11/777,712, entitled
"Producing Fluidic Waveguides", incorporated herein by reference in
its entirety. Excitation light could have any appropriate
wavelength, such as 266 nm, for example. The distance from surface
660 to obtain appropriate homogeneity can be determined, as
described, for example, in U.S. Patent Application Publication No.
2008/0013877, incorporated herein by reference; the distance can
also be sufficient to allow integration of blocking material
622.
[0141] In the illustrated implementation, the end of channel
portion 602 at right in FIG. 11 is open, providing an additional
port 666 through which fluid can enter into or exit out of channel
portion 602. Alternatively, article 600, instead of ending at
transverse end-surface 668, could extend to another area with ports
similar to ports 608, such as with a part symmetrical about the
position of surface 668; in this case, fluid could flow through
channel portion 602 between ports 608 and similar ports at the
opposite end of channel portion 602.
[0142] In the implementation in FIG. 11, the filters within filter
assembly 610 are shown in cross section, and, in this
implementation, the filters do not overlap, but rather are adjacent
to each other. They could, for example, be integrated into a recess
in the lower surface of component 630 such that they are even with
the surrounding surface of component 630 or they could be
surrounded on all sides by a layer of shadow (light blocking) or
transparent material of the same thickness; in either of these
approaches, the filters could be implemented so that there is no
step at the edges of assembly 610. The size of the gap, if any,
between adjacent filters depends, for example, on the resolution of
the technique used to produce the filters. If the filters are
produced by printing two different light-absorbing materials that
have different absorption spectra (in which case a surrounding
layer of shadow or transparent material could also be printed
around them), the registration and gaps between filters depend on
the resolution of the printing technique used; examples of such
techniques are described in U.S. Patent Application Publication No.
2007/0172969, entitled "Additive Printed Mask Process and
Structures Produced Thereby", and in co-pending U.S. patent
application Ser. No. 11/755,717, entitled "Surface Energy Control
Methods for Color Filter Printing", each of which is incorporated
herein by reference in its entirety. In general, however, the
techniques described herein do not require highly precise
positioning of filters--a small gap between filters should not
significantly affect time-varying signals that result from an
object traveling past such filters while it emanates light.
[0143] The upper part of FIG. 11 includes two graphs illustrating
intensities detected by photosensor 670 in response to two types of
objects, one emanating light of color "A", the other emanating
light of color "B". Filters 612, 616, and 620 have bands that allow
light of color "A" to pass, while filters 614 and 618 have bands
that allow light of color "B" to pass.
[0144] Curve 672 illustrates intensities indicated by sensing
results from photosensor 670 if object 640 emanates light of color
"A" as it travels along the path through channel portion 602. In
other words, the emanating light's photon energy distribution
matches the band for filters 612, 616, and 620 so that curve 672 is
high along those filters but low along filters 614 and 618; its
high value is indicated on the vertical axis as "ImaskA".
[0145] Curve 674, on the other hand, illustrates intensity
indicated by sensing results from photosensor 670 when object 640
emanates light of color "B" as it travels along the path. In this
case, the emanating light has a photon energy distribution that
matches the band for filters 614 and 618 but not for filters 612,
616, and 620, so that curve 674 is at a high intensity along
filters 614 and 618, "ImaskB", and at a low intensity
elsewhere.
[0146] Curves 672 and 674 illustrate an example in which two
different types of objects provide signals that are approximately
complementary, except at the far left along blocking material 622
where both curves are at approximately zero intensity. In a simple
implementation, for example, filters 612, 616, and 620 could be red
band pass filters, filters 614 and 618 could be green band pass
filters, each object could either be a red fluorescing particle or
tag, i.e., emanating light of color "A", or a green fluorescing
particle or tag, i.e., emanating light of color "B". As suggested,
curves 672 and 674 could be plotted based on the x-direction
position of object 640 or based on the t-position within the time
varying output signal from photosensor 670, which could be provided
continuously or by any suitable form of sampling, such as by
periodic readout at an appropriate frequency. The high intensities
of curves 672 and 674 would be reduced to the extent that blocking
material 622 prevents light from reaching photosensor 670.
[0147] As a result, output signals from photosensor 670 can be used
to distinguish types of objects, in this case to distinguish
objects that emanate light of color "A" from objects that emanate
light of color "B", and examples of techniques that distinguish
types of objects in various ways are mentioned below in relation to
exemplary implementations. In some examples, emanating light
encoded by a filter assembly with stripes of random lengths can be
analyzed by comparing a resulting time-varying signal with one or
more templates or other signals to determine an object's type,
displacement, and position to a high level of precision.
[0148] FIG. 12 illustrates two alternative implementations similar
to those in FIGS. 9-10, and with the same reference numerals, but
with filter assembly 610 on a photosensitive surface of photosensor
648. These implementations could be implemented by printing or
otherwise depositing and patterning filters 612, 614, 616, 618, and
620 and blocking material 622, such as in the manner described
above, or by producing a longitudinal sequence of band pass filters
in any other appropriate way, with some possible techniques being
described below in relation to other exemplary implementations. In
the implementation at left, photosensor 648 also operates as one
side of channel portion 602, replacing light-transmissive component
630 along at least a portion of the channel. In other words, filter
assembly 610 is positioned similarly to filter assembly 642 in FIG.
10, allowing a very small MFS. In the implementation at right in
FIG. 12, photosensor 648 is outside of channel portion 602
separated from the outer surface of component 630 by a small gap of
height g as shown. In this implementation, filter assembly 610 is
positioned similarly to filter assembly 644 in FIG. 10, but not
directly on the outer surface of component 630, so that a larger
MFS is necessary. The gap between component 630 and photosensor 648
can be maintained by spacers or other appropriate support
components, and can be sufficiently large that photosensor 648 does
not interfere with anti-resonant waveguiding within channel portion
602, which can be implemented, for example, in the ways described
in co-pending U.S. patent application Ser. No. 11/316,660, entitled
"Providing Light to Channels or Portions", incorporated herein by
reference in its entirety.
[0149] Absorption filters as described above in relation to FIGS.
9-12 can be implemented in a multitude of ways. For example, rather
than only two types of band pass filters that have bands for
respective colors, three or more types of filters with three or
more respective colors could be used. Similarly, a filter assembly
can include band pass filters and other types of absorption filters
as would be found in a shadow mask. Furthermore, with printed
filters as described above or with other filters produced with
layers of material, overlapping band pass filters could be
produced, providing additional information. In addition, absorption
filters could be combined with reflection filters, as described
below in relation to some exemplary implementations.
[0150] Filter assembly 700 in FIG. 13 illustrates some of these
variations. In the illustrated assembly, each stripe is labeled
with a description of its filter criterion. Stripe 702 is a red
band pass filter; stripe 704 is a closed filter, meaning that it
allows no transmission; stripe 706 is an open filter, meaning that
it allows full transmission; stripe 708 is a gray filter, meaning
that it passes all photon energies across a range of interest, but
at an intensity in between an open filter and a closed filter;
stripe 710 is a green band pass filter; stripe 712 is a combined
band pass filter that passes only the intersection of blue and
green; and stripe 714 is a blue band pass filter. In addition, as
can be seen, the widths of the stripes are random rather than
periodic.
[0151] The cross section in FIG. 14 illustrates one way of
implementing filter assembly 700 in FIG. 13, illustratively using
patterned layers of light absorbing material to produce different
types of filters. The implementation in FIG. 14 could, for example,
be implemented by printing or otherwise depositing and patterning
layers of material as described above.
[0152] In the cross section at the top of FIG. 14, filter assembly
700 includes red layer part 720, black layer part 722 overlapping
layer part 720, gray layer part 724, green layer part 726, and blue
layer part 728 overlapping layer part 726. Where overlaps occur,
the result is the intersection of two absorption filters: the
intersection of layer parts 720 and 722 is a closed filter, while
the intersection of layer parts 726 and 728 is a filter with a band
that is the intersection of the bands of the green and blue
filters.
[0153] The three graphs below the cross section show expected
intensity signals similar to those in the graphs in FIG. 11. Curve
730 would be for a red fluorescing particle or tag; curve 732 would
be for a green fluorescing particle or tag; and curve 734 would be
for an example where object 640 is tagged both with a red and a
green fluorescing particle so that curve 734 is a scaled sum of
curves 730 and 732. More generally, the technique of FIGS. 13 and
14 would make it possible to distinguish not only red, green, and
blue particles and tags, but also objects tagged with combinations
such as red and green, green and blue, red and blue, and red and
green and blue. Each combination results in a distinguishable time
varying signal that can be analyzed to obtain information about the
color or colors that are emanating.
[0154] Although the intensity signals described above in relation
to FIGS. 11 and 14 could be obtained from sensing results of a
single, large area photosensor, it would also be possible to use an
IC with an array of photosensing cells or an array of discrete
photosensors, in either case appropriately positioned along a path
traveled by objects past one or more filter assemblies. If an array
is used, and each element of the array is covered with a different
filter assembly, it may be possible to distinguish many different
types of particles concurrently. The number of particles to be
distinguished can be much larger than the number of elements in the
array, since each measurable distinguishing feature can provide one
axis in a principal component analysis, and multiple particles can
be distinguished along each such axis. Additional techniques that
can be used to track and distinguish objects are described in
co-pending U.S. patent application Ser. No. 11/702,328, entitled
"Distinguishing Objects", incorporated herein by reference in its
entirety. Objects can be distinguished, for example, from their
environment or background or from objects of other types; an
operation "distinguishes" objects if the operation locates,
selects, sorts, counts, or otherwise identifies an object or
controls or directs an object according to type or separates
objects or otherwise treats objects differently in some way.
[0155] Band pass filters of other types can also be used to
implement filter assemblies as described in some of the exemplary
implementations herein. For example, interference based filters can
have different bands similar to the bands described above in
relation to FIGS. 9-14.
[0156] Filter assembly 750 in FIG. 15 illustrates an implementation
in which a thin layer of transparent material creates Fabry-Perot
oscillations, and can be structured to obtain high
thickness-dependent index contrast. Assembly 750 includes filters
752, 754, 756, 758, and 760, each of which has substantially
constant thickness, but with the thicknesses of filters 752, 756,
and 760 being approximately equal to each other while the
thicknesses of filters 754 and 758 are approximately equal to each
other but smaller. Assembly 750 could be produced, for example, by
etching a deposited layer of transparent material or by imprinting
a non-solid layer of such material before it solidifies.
[0157] To the right of the cross section of assembly 750 is a graph
showing an intensity-energy function of its transmitted light. In
other words, curves 770, 772, and 774 are approximately the same
because filters 752, 756, and 760 have approximately the same
thickness. On the other hand, curves 774 and 776 are also similar
to each other but different than the others, because the
thicknesses of filters 754 and 758 are the same as each other but
different than the others. As a result, an object traveling along a
path past assembly 750 results in a time-varying signal with
changing intensity-energy function. The total transmission at each
position will relate to the overlap of the cavity's transmission
lines and the particle spectrum. The other part of the emanating
light would be reflected from assembly 750, and could also be
detected to obtain confirming information. For example, assembly
750 could be on one cover slide of a channel, and two photosensors
(not shown) could be positioned, one on the side of assembly 750
away from the channel and the other on the opposite side of the
channel to obtain sensing results for the reflected emanating
light.
[0158] FIGS. 16 and 17 illustrate two ways in which Fabry-Perot
interference-based filters could be structured to obtain band pass
filters more nearly similar to those of FIGS. 9-12. In each
implementation, optical thickness of the filter's cavity varies in
the x-direction, but the variation in optical thickness is produced
in two different ways. The general strategy in FIGS. 16 and 17 is
to provide regions that operate as band pass Fabry-Perot filters,
with different sets of filters having transmission peaks at
different photon energies. For example, one set of filters could
have a transmission peak at approximately 822 nm, while another
could have a transmission peak at approximately 833 nm, and the two
sets could have a periodic pattern as shown or any other
appropriate pattern.
[0159] Filter assembly 800 in FIG. 16 includes homogeneous bottom
distributed Bragg mirror (DBR) 802, cavity 804, and upper DBR 806.
Such a filter assembly could be produced by using techniques
described in U.S. Pat. No. 7,315,667, entitled "Propagating Light
to be Sensed", incorporated herein by reference in its entirety. As
can be see in FIG. 16, however, the optical thickness of cavity 804
has been modified by changing between two thicknesses, one larger
and one smaller, so that assembly 800 effectively includes two sets
of filters: reference numerals 810, 812, and 814 indicate three
filters with the larger thickness while regions 816 and 818 are
filters with the smaller thickness, therefore transmitting a
shorter wavelength than the filters in regions 810, 812, and 814.
The variations in thickness of cavity 804 can be produced, for
example, by etching the layer in which cavity 804 is formed after
it is deposited and before the series of layers in DBR 806 are
deposited. Alternatively, a half-tone mask could be used during
growth of cavity 804.
[0160] Filter assembly 820 in FIG. 17 similarly includes lower DBR
822, cavity 824, and upper DBR 826, each of which illustratively
has approximately uniform thickness, but with cavity 824 having
optical thickness that varies in the x-direction. As a result,
regions 830, 832, and 834 transmit a different photon energy than
regions 836 and 838. More specifically, the refractive index of
cavity regions 840, 842, and 844 is different than the refractive
index of regions 846 and 848. Differences in refractive index could
be produced in a wide variety of ways. Implantation or ion
diffusion (as in ion exchange) could be performed as is done in
fabricating waveguides for integrated optics; another approach
would be implantation-induced intermixing of multiple quantum well
(MQW) structures as in laser diode fabrication; further,
ultraviolet light-induced changes in refractive index could be used
as with germanium-doped glass used in fabricating fiber Bragg
gratings (FBG) in glass fibers; in principle, any technique that
can modify refractive index by implantation, heat, light, or other
operation could be used.
[0161] FIG. 18 illustrates an additional technique that could be
used together with the technique of FIG. 15 and possibly the
techniques of FIGS. 16 and 17. Filter component 860 includes a
wedge-shaped layer of transparent material as in FIG. 15 or
Fabry-Perot filter as in FIGS. 16 and 17, but with filter
assemblies 864, 866, and 868 formed at its upper surface such as by
techniques described in relation to FIG. 15, 16, or 17. In other
words, in addition to having filters of the types described above,
there is also a continuously varying thickness across component 860
so that, in addition to the time-varying effects of each filter
assembly, additional spectral information is contained in the
encoded emanating light, and can be obtained by appropriate
processing. With techniques such as this, it may be possible to
measure the entire spectrum with a loss of not more than 50%
(assuming full modulation) of the light, which would be
advantageous in comparison with conventional linear variable filter
approaches.
[0162] In implementations as in FIGS. 9-12, laminar flow can be
used to provide substantially uniform object speed past a filter
arrangement. In contrast, FIGS. 19-21 illustrate examples in which
laminar flow can produce non-uniform displacement or can be
modified in other ways.
[0163] FIG. 19, taken along a line similar to line 19-19 in FIG.
12, shows wall-like parts 870 and 872 with linearly decreasing
distance between them. As a result, as object 640 passes along
two-color filter assembly 610 (with filter elements 612, 614, 616,
618, and 620 illustratively periodic rather than random as in FIG.
12), its velocity increases linearly as indicated by curve 874,
either as a function of position or of time. Therefore, rather than
a periodic time-varying signal, the resulting time-varying signal
is chirped, meaning that the periods decrease linearly due to
change in velocity of object 640 due to change in the flow speed of
fluid in the channel resulting from the changing channel
dimensions. Curve 874 illustrates the resulting chirped signal,
which has intensity I(A) during regions 612, 616, and 620, and
intensity I(B) during regions 614 and 618. As can be seen, the
duration of the signal during each successive region is shorter
than the preceding region, resulting in the chirped pattern. For
the sake of illustration, the linear decrease in transition time is
exaggerated in curve 874 in comparison to the narrowing of the
channel.
[0164] The technique in FIG. 19 is only one of a variety of ways of
producing a chirped time-varying signal, and various other
techniques could be used. For example, more complex flow speed
distributions could be obtained by modifying the channel walls in
other ways or by providing devices that change the flow speed or
flow pattern within the channel, any of which would produce more
complex time-varying signals from different objects.
[0165] FIG. 20 illustrates, on the other hand, how relatively
simple time-varying signals could be produced using more
complicated techniques. In general, such techniques assume that
geometry of a channel directs flow of object 640 in a defined
manner such as periodic, chirped, or random, past a sequence of
filter elements. This allows redirection of particle flow past a
simpler filter assembly geometry, and may be advantageous in cases
where it is easier to redirect particle flow to produce a desired
time-variation of emanating light than it would be to produce a
filter assembly to produce the same time variation; for example, it
might be easier to change channel wall shapes than to produce a
desired filter assembly. In other cases, on the other hand, it
might be advantageous to obtain more abrupt or rapid signal
transitions with a well-defined filter assembly. In addition to the
techniques described below, which involve shaping or moving walls,
an object's flow within a channel could also be redirected by other
techniques; an electrically charged object such as a particle, for
example, could be redirected by electrical field variations. In
general, however, the Reynolds number in typical microfluidic and
nanofluidic implementations are so small that laminar flow
conditions are, as a practical matter, always present.
[0166] In the example in FIG. 20, wall-like parts 880 and 882 are
parallel but each of them is shaped like a sinusoidal wave,
resulting in a sinusoidal flow pattern in the channel between them.
Filter elements 884 and 886 are homogeneous of two different
colors, illustratively labeled "A" and "B". As object 640 follows
sinusoidal path 888, it moves back and forth between elements 884
and 886, passing through a small gap between them twice during each
period. Curves 890 and 892 illustrate exemplary time-varying
signals that could result from an object traveling along path 888.
Curve 890 illustrates an example of an object of a type with a
spectrum similar to color A but different from color B, while curve
862 illustrates an example of an object of a type with a spectrum
similar to color B and different from color A. As a result, the
curves are somewhat complementary, although each curve goes to
approximately 0 while path 888 is crossing stripe 894 of blocking
material between elements 884 and 886. Blocking material could also
be provided outside elements 884 and 886.
[0167] Wall-like parts 900 and 902 in FIG. 21 are substantially
straight and parallel, with filter elements 904 and 906 between
them, similar to elements 884 and 886 in FIG. 21. Motion device 908
produces relative movement between the path of object 640 and
stripe-like elements 904 and 906, as indicated by bi-directional
arrow 910. Control circuitry 912 provides signals to control
operation of motion device 908, which need not be periodic, but
could take any appropriate pattern, resulting in arbitrary
time-varying signals with features indicating different types of
objects. An alternative would be to move elements 904 and 906; more
generally, any combination of relative movements between walls 900
and 902 on the one hand and elements 904 and 906 on the other could
produce movement as indicated by bi-directional arrow 910.
Furthermore, additional variations could be produced by changing
fluid flow within the channel so that the speed or other
displacement of object 640 changes as a function of time relative
to the other movements. Motion device 908 could be set up to
produce variations in response to trigger signals indicating
incoming objects.
[0168] Curve 914 illustrates movement of object 640 between element
904, labeled "Band A", and element 906, labeled "Band B". As
illustrated, object 640 spends different lengths of time in each
region and can spend a random amount of time in each region,
resulting in a random excitation pattern. Curves 916 and 918
illustrate exemplary time-varying signals that could be produced by
the technique of FIG. 21. One type of object has a spectrum more
similar to color A of element 904, as illustrated by curve 916,
while the other has a spectrum more similar to color B of element
906, as illustrated by curve 918. As each object travels between
elements 904 and 906, it passes over stripe 919 of blocking
material between them, resulting in a brief interruption of the
emanating light, so that each curve goes briefly to 0. In curve
916, the intensity along element 904 is I(A1), while the intensity
along element 906 is I(B1), a lower value linearly. Conversely,
curve 918 illustrates that the intensity is higher along element
906, at intensity I(B2), and lower along element 904, at intensity
I(A2). The two curves are, in general, complementary, except for
times when they are passing stripe 919 between element 904 and 906;
object 640 can be moved instantaneously between Band A and Band B,
moving very quickly across stripe 919, so that the time in which it
is passing stripe 919 are very brief.
[0169] FIGS. 22-24 illustrate implementations of filter
arrangements in which filter assemblies are on opposite sides of
channel 920. In the illustrated implementation, detector 922, shown
on the near side of channel 920, includes one filter assembly,
while detector 924, on the far side of channel 920, includes
another filter assembly. Although each detector could be
implemented in a wide variety of different ways, to obtain
information about emanating light and objects from which light
emanates, FIGS. 23 and 24 illustrate an example in which detector
922 includes a periodic filter assembly with periodicity in a
direction transverse to channel 920, labeled the y-direction, and
detector 924 includes a random two-color filter assembly with a
longitudinal sequence in the x-direction, though other angles
between the x- and y-directions might also be useful including, in
some cases, implementations in which they are parallel. In the
illustrated case, sensing results from detector 922 include signals
modulated in the y-direction, while sensing results from detector
924 indicate signals modulated in the x-direction. The two
modulations can be used to obtain information about an object from
which light is emanating.
[0170] As shown in FIG. 23, detector 922 can be implemented with
photosensor 926 on a photosensitive surface of which are filters
927, periodic in the y-direction; each of filters 927 is
illustratively a red band pass filter, but they could instead be
any other color or closed filters or intermediate intensity gray
scale filters, and could be implemented with absorption,
reflection, or interference-based filtering techniques as described
above. Similarly, FIG. 24 shows an implementation of detector 924
in which photosensor 928 has filter assembly 610 (FIG. 9) on its
photosensitive surface; photosensor 928 could also have a periodic
filter superimposed on filter assembly 624 or in place of filter
assembly 624, in which case it might include green filters (not
shown).
[0171] A wide variety of other arrangements similar to FIGS. 22-24
would be possible, including, for example, another type of template
layer on one side of channel 920 to provide a desired signal as
described in co-pending U.S. patent application Ser. No. 12/022,485
entitled "Obtaining Information from Time Variation of Sensing
Results", incorporated herein by reference in its entirety, and a
periodic mask layer to provide a periodic signal on the other side
of channel 920; in this implementation, the periodic signal could
be constantly analyzed to obtain values indicating displacement of
an object currently flowing through channel 920, which could be
used to determine an appropriate time scale for correlation with
the template signal similar to techniques described. In another
possible variation, emanating light from fluorescence could be
photosensed on one side of channel 920 and emanating light due to
scattering, for example, could be photosensed on the other
side.
[0172] Some of the exemplary implementations described below
involve filter assemblies that combine periodic signals additively
with template signals from filter sequences similar to some of
those described above. The resulting time-varying signal emerges
from the filter assembly with two different spatially varying
patterns imposed on it. To produce such a signal, for example, a
radial sequence or "stack" of filters similar to that shown in FIG.
4 could be used. Within a stack of filters, for example, one layer
could be a template layer with an appropriate pattern to produce
the template signal, while another layer could be a periodic layer
with an appropriate pattern to produce the periodic signal; each of
the template layer and periodic layer could have rectangles or
other closed polygons of zero opacity surrounded by regions with
opacity 0.5.
[0173] FIGS. 25 and 26 illustrate an alternative approach that can
be used with reflective gray scale filters, producing a single
filter assembly equivalent to a desired radial sequence or stack of
filters. To obtain filters as in FIGS. 25 and 26, thickness
definitions of two filter layers can be overlaid using software
tools and the thicknesses of overlapping regions can be added,
resulting in regions with thicknesses of 0, 0.5, and 1 in the
example given above; the two filter layers could both be oriented
with variation in the same direction as in FIGS. 25 and 26, similar
to the techniques of FIGS. 9 and 13, or could be oriented with
variation in different directions, e.g. orthogonal to each other.
For implementations in which layer thickness does not appropriately
define or determine the desired equivalent filters structure or its
optical variation, the techniques in FIGS. 25 and 26 could be
modified to first overlay optical feature definitions of the
filters in which regions have defined optical feature values that
determine the desired variation, thus obtaining an optical feature
definition of the desired equivalent filter; the optical feature
definition could then be converted to a layout-type description of
the equivalent filter in which each region has a defined optical
thickness or other characteristic that can be produced to provide
the region's value for the optical feature.
[0174] The techniques of FIG. 25-26 take advantage of the fact
that, in general, superpositions of filters are commutative, in the
sense that the resulting transmission or reflection function is the
same regardless of the order in which filters are superimposed.
There are, of course, exceptions, such as where interference
effects can occur if filters are in a specific order, or where
alignment or other relationship of filter features can result in
loss of different information depending on the order of the
filters.
[0175] If the equivalent filter definition is a thickness
definition to produce a purely transmissive/reflective filter with
no color variation, and if partial etching can be performed, an
equivalent filter that approximates the equivalent filter
definition can be constructed by first depositing a highly
reflective material, such as chromium, over the entire filter
assembly, and by then partially etching the reflective material
away in regions with thickness 0 or 0.5 to an appropriate extent,
leaving a thin, partially transmitting layer, after which the
remaining reflective material can be etched away in regions with
thickness of 0. Where partial etching is unreliable, other
techniques may be used, such as by techniques that deposit a first
patterned layer of thickness 0.5 with any suitable patterning
technique, then depositing over it a second patterned layer of
thickness 0.5 that is patterned without etching, such as with
liftoff or other patterning techniques that do not require etching.
Furthermore, similar techniques might be applied to produce layered
filter structures that include DBRs of varying
transmission/reflectivity and/or cavities of varying optical
thickness, such as those described above in relation to FIGS.
16-18; variation in cavity thickness could result from any
appropriate combination of thickness variation and refractive index
variation, produced with any appropriate techniques.
[0176] Filter 930 in FIG. 25 is equivalent to the combination of a
random filter and a periodic filter, superimposed one on the other.
Curve 932 shows the shape of the random filter, while curve 934
shows the shape of the periodic filter; as can be seen, the random
and periodic filters both have only two thickness levels, either 0
or 0.5, but filter assembly 930 has three thickness levels,
corresponding to 0, 0.5, and 1. Curve 936 shows a resulting
transmission function. Emanating light passing through filter
assembly 930 includes both displacement and position information
about an object from which it emanates, and allows time-scaling
techniques to extract that information, as described below.
[0177] The technique illustrated in FIG. 25 can be adjusted as
suggested by dashed lines 938 within filter 930. In other words,
total light output can be changed by scaling the amplitude of the
thickness levels: rather than 0, 0.5, and 1, for example, thickness
levels of 0, 0.2, and 0.4 could be used, allowing greater light
transmission. It may be necessary, however, to make a tradeoff
between greater light output, and therefore total signal intensity,
on the one hand, and greater light modulation on the other--greater
light modulation may facilitate calculation of displacement and
position within a given observation region. The mask suggested by
dashed lines 938 emphasizes total light output because it has
reduced thickness and, conversely, increased transmission, with a
thickness of 0 being equivalent to transmission of 1 and vice
versa. The scaling suggested by dashed lines 938 may require great
precision: the x-direction scale of features in assembly 900 may be
as great as 10 .mu.m, while a useful thickness may be as thin as 10
nm of chromium.
[0178] Similarly, filter assembly 940 in FIG. 26 is equivalent to
the combination of a chirp filter represented by curve 942 and a
periodic filter represented by curve 944. A combination of chirp
and periodic filters can make it possible to more efficiently
extract displacement and position information about objects that
may have different speeds. Curve 946 shows a resulting transmission
function, which allows information extraction.
[0179] A stack-equivalent filter assembly as in FIGS. 25 and 26 can
in some cases have a smaller MFS than either of the simpler
non-uniform filters. As mentioned above, loss of resolution can
occur for light emanating from objects approximately as large as
the MFS.
[0180] FIG. 27 illustrates one way in which a longitudinal sequence
of filters, such as a random band pass filter arrangement as
described above in relation to FIGS. 9-12 can be combined with a
reflective gray scale filter arrangement, illustratively a periodic
gray scale filter. Filter arrangement 950 in FIG. 27 includes
filter subassembly 952 with a longitudinal sequence similar to that
described above in relation to FIGS. 9-12. On the upper surface of
subassembly 952 is a periodic filter subassembly with regions 954,
each having an intermediate transmission level such as 0.5. As a
result, filter assembly 950 combines the technique of FIG. 25 with
that of FIG. 12, providing distinguishable time-varying signals for
emanating light of different colors, and also modulating the
emanating light to allow time-scaling techniques as described
below. In effect, the time-scaling operations can be performed in
the same way for each emanating color's signal, and the different
color signals can be used to distinguish types of objects after
time scaling.
[0181] The flow chart in FIG. 28 illustrates ways in which
information about objects can be obtained and used by CPU 402 (FIG.
6); the technique of FIG. 28 illustratively extracts information
such as a type, a position, or a spectral difference, and uses such
information to distinguish objects. FIG. 28 also suggests ways in
which routines 440, 442, and 444 (FIGS. 6 and 7) could be
implemented. Although suitable for CPU 402, operations in FIG. 28
could be implemented with a wide variety of different types of
circuitry with or without a CPU. Furthermore, although described in
terms of time-varying signals from photosensors, the technique of
FIG. 28 could be applied to any time-varying sensed signals,
including, for example, capacitively sensed signals from charged
particles with encoded information due to shapes, sizes, and
positions of electrodes.
[0182] The operation in box 970 obtains one or more encoded
time-varying signals from a photosensor arrangement as one or more
objects travel along respective paths past a filter arrangement.
The technique could be implemented with a single photosensor along
the paths, but it might also be possible to implement with two
photosensors on opposite sides of the paths or with other
photosensor arrangements. The objects can, for example, travel
through a channel as described above in relation to FIGS. 8-12 and
19-21 and the time-varying signals can be encoded in any of a wide
variety of ways using filter arrangements, including one or more of
those described above, with or without displacement control and/or
spatially modulated excitation; excitation techniques that could be
used are described in co-pending U.S. patent application Ser. No.
12/023,436 entitled "Producing Time Variation in Emanating Light",
also incorporated herein by reference in its entirety. For example,
if one of the filter arrangements is at least partially
non-periodic or if displacement control or excitation is at least
partially non-periodic, a respective template of the resulting
non-periodic pattern for each of a number of types of objects can
be used to perform a correlation operation; in other
implementations, two differently encoded time-varying signals can
be obtained in box 970 and correlated with each other. Note,
however, that two types could be distinguished based on a single
template, especially if their time-varying signals are sufficiently
complementary that one results in correlation and the other in
anti-correlation with the template.
[0183] The operation in box 970 can include providing any
appropriate control signals to other components of the system,
including signals to read out sensing results of photosensors. The
control signals could be implemented as in routines 440 and 442
(FIG. 6), with CPU 402 providing signals through device I/O 420 to
one or more of devices 422 through 424. For example, fluid flow
speed could be adjusted and channel wall movement could be
controlled as described above in relation to FIG. 21. In order to
obtain the time-varying signals, CPU 402 could provide signals
through IC I/O 410 to obtain photosensed quantities from ICs 412
through 414.
[0184] The operation in box 972 performs a correlation or other
comparing operation on one or more time-varying signals from box
970, such as comparing two encoded signals with each other or
comparing one encoded signal with a respective template of a
non-periodic encoding pattern for each distinguishable type of
object. As used herein, the term "correlation operation"
encompasses any of a variety of mathematical operations that can be
performed on a pair of time-varying functions, with or without
scaling, and that obtains a similarity measure as a function of
time-alignment. This correlation operation can be implemented, for
example, as described in co-pending U.S. patent application Ser.
No. 11/698,338, entitled "Method and System for Evaluation of
Signals Received from Spatially Modulated Excitation and Emission
to Accurately Determine Particle Positions and Distances",
incorporated herein by reference in its entirety. Additional
correlation and other comparison techniques that could be used are
described in co-pending U.S. patent application Ser. No. 12/022,485
entitled "Obtaining Information from Time Variation of Sensing
Results", also incorporated herein by reference in its
entirety.
[0185] A correlation operation in box 972 can produce correlation
results for each pair of waveforms that is compared. For example,
if box 972 compares an encoded time-varying signal from box 970
with each of N templates for N types of objects, N correlation
results are produced.
[0186] The graphed curves in box 974 illustrate two types of
correlation results: The upper curve illustrates a correlation
result where two time-varying waveforms are correlated, i.e. highly
similar at the time alignment designated t.sub.p; the lower curve
illustrates a correlation result where two time-varying waveforms
are anti-correlated, i.e. highly dissimilar at the time alignment
designated t.sub.p'. In each case there is a peak, with the peak in
the correlated case marked to show its amplitude A and with the
anti-correlated case having an inverted peak of similar amplitude.
If correlation is performed on a continuous basis, correlation
results could similarly be continuously obtained for each template
with which comparison is made, with each object's travel past the
filter arrangement producing a peak, an inverted peak, or a feature
in between the two for each template.
[0187] The operation in box 980 obtains a time-varying waveform
that equals or approximates the time derivative d/dt of each
correlation result from box 972. For the correlated case, a
derivative waveform like the graphed curve in box 982 is obtained,
with a positive peak followed by a negative peak, with a zero
crossing at t.sub.p, and with the contrast or differential quantity
between the peaks again being the amplitude A. For the
anti-correlated case, a derivative waveform like the graphed curve
in box 984 is obtained, with a negative peak followed by a positive
peak, with a zero crossing at t.sub.p', and with the contrast or
differential quantity between the peaks being amplitude A', the
amplitude of the inverted peak in the lower graph in box 974. The
amplitudes obtained in this manner are, in general, free of
offsets, allowing direct comparison to obtain spectral
information.
[0188] The operation in box 986 uses derivative waveforms from box
980 to extract information for objects passing the photosensor. The
extracted information could, for example, be a type based on
whether an object resulted in correlation, anti-correlation, or
neither with a given template; position based on the time at which
a zero crossing occurs in correlation or anti-correlation; and
spectral difference, e.g. a difference of emission, absorption, or
scattering spectrum, based on the amplitude or contrast between
positive and negative peaks from correlation and anti-correlation,
respectively. Features of a derivative waveform could be found and
measured using various techniques. The operation in box 988 can
then be performed to distinguish objects using information
extracted in box 986, such as by obtaining counts of different
types of objects or ratios between such counts, or with other
operations as described above in relation to FIG. 7.
[0189] The operations in boxes 972, 980, and 986 could be
implemented, for example, as parts of one or both of routines 442
and 444 (FIG. 6). The operation in box 988 could be implemented as
part of routine 444. In general, these operations could be
implemented to handle signals from each object separately or to
handle a signal received concurrently or in series from a number of
objects, in which case minimum differences, such as in positions or
speeds, may be necessary to allow separation of signals from
different objects. Any appropriate combination of serial and
parallel operations could be implemented in any appropriate
circuitry. Data streams or other data structures defining waveforms
such as templates could be stored and retrieved as needed by
routines 442 and 444, such as in memory 408 (FIG. 6). Similarly,
intermediate and final results of operations in boxes 972, 980,
986, and 988 could similarly be stored and retrieved as needed.
[0190] Comparison techniques other than correlation could be
employed, but correlation techniques can be advantageous because
they are typically not sensitive to noise, such as an AC power
frequency. For example, preliminary smoothing or other
preprocessing of waveforms is typically unnecessary for
correlation, and available techniques for computing correlations
can produce useful results even with S/N ratios significantly less
than 1.0. It is, however, necessary to satisfy minimum sampling
requirements if waveforms are digitized for correlation; in
accordance with the Nyquist frequency, each waveform should be
sampled at least twice during the time duration of its minimum
feature size.
[0191] Some techniques as described above have been successfully
applied to simulated time-varying waveforms. In particular, time
scaling techniques have been found to improve S/N ratio of a
simulated observed signal that contains both an encoding based on a
template and also additive noise, and where the observed signal has
an unknown time scaling that occurs before it is observed; S/N
ratio of 0.5 has been obtained and 0.1 appears achievable. These
results could be obtained with particle speeds up to 0.5 m/sec and
higher speeds up to a few m/sec appear to be feasible, with
particles having effective sizes down to 0.6 .mu.m, and with
particle separations down to a given implementation's MFS. A
demonstration included counting CD4 in a whole blood sample; single
tag detection was shown to be feasible.
[0192] Where a simulated observed signal includes or is accompanied
by a simulated concurrent periodically modulated signal, time
scaling of a template waveform based on a scaling factor from the
periodically modulated signal has successfully produced matching
correlation results, indicating correlation or anti-correlation as
appropriate and making spectral information available, in effect
allowing concurrent detection of multiple colors with a single
detector such as a large-area photosensor. Because an object
receives different excitations at almost the same time and location
(due, for example, to interdigitated or otherwise patchworked or
patterned excitations), differences in absorption and excitation
spectra can be measured with very high precision; similarly,
because different spectral subranges of an object's emission
spectra are measured at almost the same time and location (due, for
example, to interdigitated, or otherwise patchworked or patterned
filter arrangements), differences in emission spectra can be
measured with very high precision; therefore, with one or both of
patterned excitation and patterned filtering, many types of errors
cancel out, including time-dependent factors such as bleaching,
intermixing, diffusion and also errors induced by excitation
differences such as temperature gradients and optical
misalignments. Particle position can be precisely determined from
fine structure of correlation results. As noted above, simulation
results show that spatial resolution of less than 1.0 .mu.m is
possible, and single fluorescence markers can be detected, making
detection possible with smaller amounts of consumables such as
markers. The techniques appear appropriate for native fluorescence,
allowing agent-less detection.
[0193] Some of the implementations described above in relation to
FIGS. 1-28 are examples of a method of using a filter arrangement.
While an object travels along a path past the filter arrangement
and emanates light within an application's range of photon
energies, the method transmits/reflects at least some of the
emanating light through the filter arrangement. In doing so, the
method includes at least one of the following two: First, while the
object is in each of two or more segments of the path, the method
transmits/reflects respective portions of the emanating light
through respective positions of a filter assembly within the filter
arrangement; each of the respective positions has a respective
transmission function, and the transmission functions of at least
two of the positions are sufficiently different that time variation
occurs in the emanating light between at least two of the segments.
Second, while the object is in each of a series of segments of the
path, the method transmits/reflects a respective portion of the
emanating light through a filter component within the filter
arrangement, and the filter component has a combined transmission
function in which a set of simpler transmission functions is
superimposed; the set of transmission functions is superimposed
such that time variation occurs in the emanating light in
accordance with superposition of first and second simpler
non-uniform transmission functions in the set.
[0194] In specific implementations, the method can do both, i.e. it
transmits/reflects respective portions of the emanating light
through respective positions of a filter assembly and also
transmits/reflects a respective portion of the emanating light
through a filter component. The filter assembly can include first
and second filter elements that include first and second respective
positions, and each of the first and second filter elements can
have a respective transmission function that is approximately
uniform for light from its respective segment of the path. For
example, the transmission functions of the first and second filter
elements can be spectrally different, and the method can encode
spectral information in time variation of the emanating light.
Also, the transmission functions can be different in
transmitted/reflected intensity, and the method can encode
intensity information in time variation of the emanating light.
More generally, the method can encode information about the object
in time variation of the emanating light, such as information
indicating the type of the object.
[0195] In further specific implementations, each of the segments of
the path can be at least approximately as large as the object's
size. The combined transmission function can have a minimum feature
size (MFS) at least approximately as large as the object's sizes.
The series of segments in which the method transmits/reflects
emanating light through the filter component can be substantially
continuous. Similarly, the segments from which emanating light is
transmitted/reflected through respective positions of the filter
assembly can be segments within a sequence in part of the object's
path. The method can cause the object to travel along the path with
non-uniform displacement, such as by changing its displacement rate
or its displacement direction. The object can be, for example, a
biological cell or virus, and the application can be flow
cytometry.
[0196] The simpler non-uniform transmission functions that are
superimposed can include at least one that is non-periodic.
Examples of simpler transmission functions can include, in addition
to periodic functions, random and chirped functions.
[0197] Some of the implementations described above in relation to
FIGS. 1-28 are examples of apparatus that includes a fluidic
structure with a channel through which objects can travel along
respective paths during operation and an encoding component with a
filter arrangement that can receive light emanating from objects in
the channel. In response to input light emanating from an object,
the filter arrangement provides output light. The filter
arrangement includes at least one of the following two: First, a
filter assembly with a set of positions, each having a respective
transmission function; a sequence of segments of an object's path
including at least two segments from which respective positions in
the set receive emanating light, and the transmission functions of
the positions are sufficiently different from each other that time
variation occurs in the output light while the object travels
through the sequence of segments. Second, a filter component that
receives input light from a segment of an object's path and has a
combined transmission function in which a set of two or more
simpler transmission functions are superimposed; the set includes
first and second simpler non-uniform transmission functions, and
the set is superimposed such that time variation occurs in the
output light in accordance with superposition of the first and
second simpler non-uniform transmission functions while the object
travels through the segment.
[0198] In specific implementations, the apparatus can encode
information about the object in time variation of the output light.
The apparatus can also include a photosensing component that
photosenses the time-varying output light and provides sensing
results, such as with electrical signals. The apparatus can also
include a processing component that responds to the sensing
results, performing operations to obtain data indicating
information encoded in the output light. The processing component
can be programmed, for example, to perform a comparing operation to
obtain comparison results between time-varying waveforms, at least
one of which is from the sensing results, and can use the
comparison results to obtain data indicating at least one spectral
difference between the time-varying waveforms; the comparing
operation can be correlation, and the time-varying waveforms can
include a sensed time-varying waveform and a template time-varying
waveform.
[0199] In specific implementations, the filter arrangement can
include one or more of many different filter components, such as an
absorption filter; an interference-based filter; a
light-transmissive and/or light-reflective filter; a longitudinal
sequence of filters that vary in a periodic, random, or chirped
pattern; filter elements of two or more colors; filter elements of
two or more gray levels; overlapping filter elements; a
longitudinal sequence of filter elements that includes binary, gray
level, and color filter elements; two or more lengthwise extending
filter elements; a radial sequence of filter elements; two
orthogonally striped filter assemblies; filter assemblies on
opposite sides of a fluidic channel; and a stack-equivalent filter.
The filter component can include the filter assembly, which can
have one of the first and second simpler non-uniform transmission
functions. At least one of the simpler non-uniform transmission
functions can be a periodic, random, or chirp function of
position.
[0200] In specific implementations, the apparatus can be a flow
cytometer, with the objects being, for example, biological cells or
viruses. In other implementations, the apparatus can be a scanning
device.
[0201] Some of the implementations described above in relation to
FIGS. 1-28 are examples of a method that transmits/reflects light
emanating from objects through a filter arrangement while each
object travels along a respective path past the filter arrangement.
The method can transmit/reflect at least some of the object's
emanating light through a longitudinal sequence of filter elements
within the filter arrangement, while the object travels through a
segment of its path. The longitudinal sequence includes a first
subset of filter elements that have approximately a first
transmission function and a second subset that have approximately a
second function, with the first and second transmission functions
being sufficiently spectrally different from each other that time
variation occurs in the emanating light while the object travels
through the segment.
[0202] In specific implementations, the time variation encodes
spectral information about the object, and the method can
photosense the emanating light to obtain photosensing results, then
use the photosensing results to obtain data indicating the encoded
spectral information. The photosensing results can indicate sensed
time-varying waveforms, and the method can perform a comparing
operation on a set of time-varying waveforms to obtain comparison
results, with at least one of the time-varying waveforms being a
sensed time-varying waveform; the method can use the comparison
results to obtain data indicating at least one spectral difference
between the time-varying waveforms.
[0203] In further specific implementations, where objects have
different respective emanation spectra, the method can detect a
difference between the spectra. The longitudinal sequence can be a
sequence of spatially patterned filter elements that includes at
least one spatially patterned color filter element and at least one
spatially patterned non-color filter element such as a black and
white or gray level filter element. More generally, a sequence of
patterned filter elements can include patterns sufficiently
different that different information items are concurrently encoded
in the emanating light without loss of information.
[0204] Some of the implementations described above in relation to
FIGS. 1-28 are examples of a method that transmits-reflects light
from objects through a filter arrangement while the objects travel
past the filter arrangement. The method transmits-reflects at least
some of an object's emanating light through a filter component
within the filter arrangement while the object travels through a
segment of its path. The filter component has a combined
transmission function in which a set of simpler transmission
functions is superimposed. The set includes first and second
simpler non-uniform transmission functions and is superimposed such
that time variation occurs in the object's emanating light in
accordance with superposition of the first and second simpler
non-uniform transmission functions while the object travels through
the segment.
[0205] In specific implementations, at least one of the first and
second simpler non-uniform transmission functions is a
spectrally-dependent function that transmits more than one color as
a function of position. The first non-uniform transmission function
can be a non-periodic spectrally-dependent transmission function
and the second can be a period transmission function. At least one
of the first and second non-uniform transmission functions can be a
spectrally-independent transmission function that transmits black
and white and/or gray scales. At least one of the simpler
non-uniform transmission functions can be periodic, random, or
chirped. As above, the time variation can encode information, and
photosensing results can be used to obtain data indicating the
encoded information.
[0206] Implementations as described above in relation to FIGS. 1-28
could be advantageously applied in a wide variety of sensing
applications, possibly including, for example, fluorescence- or
impedance-based flow cytometry or other biodetector applications
that seek a signature of a particle of unknown velocity; such
biodetectors often use microfluidic channels with inhomogeneous
flow profiles, causing variation in particle velocity. The
techniques can be used to count or obtain ratios between
fluorescing objects of different types, such as different types of
tagged cells, particles, tagged DNA, and so forth. In such an
application, calibration can be performed using known objects,
e.g., tagged beads, with known velocities to obtain template
waveforms that include deviations caused by fabrication tolerances
but can then be compared with sensed waveforms to obtain
information about unknown objects. To improve S/N, known and sensed
waveforms can be correlated, such as after time scaling of each
known waveform. If a sensed waveform includes or is accompanied by
periodic modulation, a periodicity value such as a frequency can be
used to obtain a scaling factor for time scaling before
correlation, allowing more rapid correlation than if a brute force
technique is used to find a satisfactory time scaling.
[0207] Implementations described above may be advantageous in
biodetector applications that require compact, low-cost components
without critical optics and with high sensing efficiency. Such
applications might include point-of-care flow cytometry (such as
with an integrated flow cytometer), DNA analysis, proteomics, and
so forth.
[0208] Implementations described above could successfully detect
native fluorescence differences between biological materials. Most
biological cells are composed of only a few basic building blocks
and, therefore, exhibit similar native fluorescence spectra.
Interdigitated or otherwise patchworked or patterned filter
arrangements like those above are particular suitable for
differentiation of objects based on their native fluorescence
signals because the techniques are sensitive enough to detect the
native fluorescence from a single cell and allow direct measurement
of distinguishing features such as intensity ratios in emission
spectra. In addition, implementations of the techniques can combine
advantages of excitation and emission spectroscopy in a rugged and
compact system.
[0209] Also, implementations described above could be applied in
scanning of bio-chips or documents where objects have different
emanation spectra, and so forth. The techniques may also be
applicable in various low S/N ratio systems in which a known signal
is bounced off an object traveling at an unknown velocity, such as
where object velocity is on the order of signal propagation
velocity, as in SONAR. The techniques may be especially
advantageous where precise information about position, speed, or
type of objects is sought.
[0210] Exemplary implementations described above employ
photosensors or impedance-based sensors with specific features, but
a wide variety of sensors could be used to obtain sensing results
indicating values of various parameters other than emanating light
intensity, parameters that can have time variation that indicates
information about objects. Similarly, implementations described
above involve sensing information about objects that are moving in
fluidic channels or that are moving relative to a sensor such as in
scanning, but various other types of fluidic implementations or
other implementations in which objects move in various other ways
could be sensed to obtain sensing results suitable for techniques
described above. For example, information could be obtained from
native fluorescence of particles in an air stream. Also, an
excitation pattern could be scanned across a glass slide with
immobilized analyte particles such as tagged cells or DNA spots, to
obtain emanating fluorescent light.
[0211] Components of exemplary implementations as described above
could have various shapes, dimensions, or other numerical or
qualitative characteristics other than those illustrated and
described above. Similarly, although the exemplary implementations
generally involve sensing from a single fluidic channel,
implementations could readily include multiple parallel channels,
allowing parallel sensing and readout and larger scale sensing.
[0212] Some of the above exemplary implementations involve specific
types of fluidic components, filter components, light source
components, displacement control components, sensors, and so forth,
but the invention could be implemented with a wide variety of other
types of components. For example, some implementations use specific
types of spatial modulation based on one or more of an excitation
pattern, a filter assembly, and/or displacement control, but
various other types of spatial modulation could be used, including
any appropriate combination of color, gray level, and black and
white patterning and including other patterning techniques such as
patterned sensing; for example, in a fluidic implementation, a
filter assembly or a patterned photosensor could be printed or
otherwise produced on an inward wall or other boundary of a channel
or in another appropriate location. Also, some exemplary
implementations use specific types of processing, such as digital
signals obtained after converting sensed analog values. In general,
however, the invention could be implemented with any suitable
signal processing techniques, including any appropriate combination
of analog and digital processing; either or both of two compared
waveforms could be obtained in analog or digital form, and any
combination of time scaling could be performed before comparison.
Further, some exemplary implementations use large area
photosensors, but various ICs with photosensing arrays might be
used.
[0213] Some of the above exemplary implementations involve specific
types of emanating light, e.g. fluorescence, and specific types of
excitation and filtering suitable to fluorescent light, but these
are merely exemplary. The invention could be implemented in
relation to various other types of optical signals in various other
ranges of photon energies or with any other appropriate sensed
stimuli.
[0214] Some of the above exemplary implementations involve specific
materials, such as in fluidic structures with light-transmissive
components or in filtering arrangements with reflective material or
light blocking material such as amorphous silicon, but the
invention could be implemented with a wide variety of materials and
with layered structures with various combinations of sublayers.
Thicknesses of layers may vary across any suitable range.
[0215] The exemplary implementation in FIG. 6 employs a CPU, which
could be a microprocessor or any other appropriate component.
Furthermore, as noted above, operations could be performed
digitally or with analog signals, and could be done either on the
same IC as a photosensor array, on other components, or on a
combination of the two, with any appropriate combination of
software or hardware.
[0216] The above exemplary implementations generally involve use of
encoding/sensing arrangements, sensors, photosensors, excitation
arrangements, filter arrangements, displacement control
arrangements, and so forth following particular operations, but
different operations could be performed, the order of the
operations could be modified, and additional operations could be
added within the scope of the invention. For example, readout of
sensed quantities from a sensor to obtain a sensed time-varying
waveform could be performed serially or in parallel, and, with an
array, could be performed cell-by-cell or in a streaming operation.
Principal component analysis could be applied to specifically
chosen intensity ratios in the emission spectrum in distinguishing
cells or other objects, possibly allowing identification. Multiple
photosensors along a channel could measure different intensity
ratios in the emission spectrum, possibly allowing identification
of objects based on either emission characteristics. Dyes that are
very similar may be distinguishable if they reveal only slightly
different emission spectra, and use of similar dyes could be
advantageous in satisfying pH requirements within cytometers.
[0217] While the invention has been described in conjunction with
specific exemplary implementations, it is evident to those skilled
in the art that many alternatives, modifications, and variations
will be apparent in light of the foregoing description.
Accordingly, the invention is intended to embrace all other such
alternatives, modifications, and variations that fall within the
spirit and scope of the appended claims.
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